2-polysubstituted aromatic ring-pyrimidine derivatives, preparation and medical application thereof

ABSTRACT

The present invention provides a 2-polysubstituted aromatic ring-pyrimidine derivative and an optical isomer thereof, or a pharmaceutically acceptable salt or solvate thereof, the compound, and an optical isomer thereof or a pharmaceutically thereof acceptable salts or solvates can be used in the preparation of anti-tumor drugs. The invention designs and synthesizes a series of novel small molecule Chk1 inhibitors by using N-substituted pyridin-2-aminopyrimidine obtained by structure-based virtual screening as a lead compound, and carries out Chk1 kinase inhibitory activity test. The experiment confirmed that said compounds possess potent anticancer activity, Chk1 kinase inhibitory activity, and are promising Chk1 inhibitors, and can be used as new cancer therapeutic drugs, which can be applied to treat solid tumors or hematologic tumors related to proliferative disease of human or animal. The 2-polysubstituted aromatic ring-pyrimidine derivatives provided by the present invention has the structure of the formula I:

CROSS REFERENCE OF RELATED APPLICATION

This is a U.S. National Stage under 35 U.S.C 371 of the InternationalApplication PCT/CN2017/110029, filed Nov. 8, 2017, which claims priorityunder 35 U.S.C. 119(a-d) to CN 201610988036.4, filed Nov. 10, 2016.

BACKGROUND OF THE PRESENT INVENTION Field of Invention

The present invention relates to the field of medicine, and moreparticularly to a 2-polysubstituted aromatic ring-pyrimidine derivativeand an optical isomer thereof, or a pharmaceutically acceptable salt orsolvate thereof, a pharmaceutical composition therewith and the same inthe field of antitumor applications.

Description of Related Arts

With the changes in human living environment and the aging of thepopulation, malignant tumors are seriously threatening human life. InChina, malignant tumors have become the first deadly disease.Traditional cancer treatment methods include surgery, radiation therapyand drug chemotherapy, among which drug chemotherapy is the mostimportant. In recent years, as tumor molecular targets have beengradually explained, many targeted anti-tumor drugs have enteredclinical applications, but due to the complexity of the tumors andgenetic diversity, single-targeted drugs are not enough to cure tumors.Traditional chemotherapeutic drugs are mostly DNA-damaging drugs, whichinduce tumor cell apoptosis by directly interfering with DNA synthesisof tumor cells, regulating DNA transcription and repair, and prolongingthe survival of cancer patients. However, due to its poor selectivity,it can cause a variety of toxic side effects, and it will producesignificant drug resistance during the treatment. Therefore, accordingto the action characteristics of DNA-damaging drugs, drugs with lowtoxicity are developed to be combined with DNA-damaging drugs, which canreduce the therapeutic effect of DNA-damaging drugs while reducing thetoxic side effects and more risk of drug resistance. Among them, thedevelopment of cell cycle-related drugs and their strategies incombination with DNA-damaging drugs have attracted great interest andattention from drug researchers in recent years.

Eukaryotic cells have their own regulatory mechanisms. When exposed toexternal stimuli such as radiotherapy or chemotherapy, it can betemporarily blocked in the G1, S or G2/M phase for DNA repair, and aftercompletion of the repair, it will enter the next phase. A large numberof protein kinases in cells interact with the same or differentsignaling pathways, forming an intricate signal network that regulatescell growth, proliferation, angiogenesis, metastasis, apoptosis andother life activities. Among them, the tumor gene suppressor protein p53is mainly responsible for the regulation of the G1 checkpoint, while theS and G2/M phases are mainly regulated by the cell cycle checkpointkinase 1 (Checkpoint kinase 1). Most tumor cells rely more on S or G2/Mphase arrest due to the loss of p53 function as a defense mechanism forDNA damage-induced apoptosis. In the p53-deficient tumor cells,inhibition of Chk1 protein can abrogate cell cycle arrest and directlyinduce tumor cell apoptosis, while normal cells are temporarily blockedin G1 phase due to the existence of intact p53 regulatory mechanism.Therefore, Chk1 inhibitor can be used as an adjuvant therapy toselectively enhance the sensitivity of tumor cells to radiotherapy orchemotherapy and improve the therapeutic effect.

In addition, in the context of specific genetic defects, such as theinherent DNA damage is too high to cause large replication pressure,Chk1 inhibitors can also be used alone, killing tumor cells through a“synthetic lethal” mechanism to achieve therapeutic purpose. Based onthis therapeutic strategy, Chk1 inhibitors can be used alone in thetreatment of B-cell lymphoma, leukemia, neuroblastoma, and somesensitive tumors with high expression of proto-oncogenes such as breastand lung cancer.

In the past two decades, small molecular compounds of differentstructural types have been discovered as Chk1 inhibitors, and thesecompounds have shown strong anti-tumor effects in preclinicalevaluation. At present, 11 small molecule Chk1 inhibitors have enteredclinical research, which proves the correctness of Chk1 as a tumortreatment target.

SUMMARY OF THE PRESENT INVENTION

An object of the present invention is to provide a 2-polysubstitutedaromatic ring-pyrimidine derivative and an optical isomer thereof, or apharmaceutically acceptable salt or solvate thereof. It is a novel2-polysubstituted aromatic ring-pyrimidine derivative with stronganticancer activity and Chk1 inhibition.

The 2-polysubstituted aromatic ring-pyrimidine derivatives provided bythe present invention comprise the structure of the general formula (I):

and an optical isomer thereof or a pharmaceutically acceptable salt orsolvate thereof;

wherein ring A is selected from substituted or unsubstituted five- orsix-membered aryl groups, and contains from 1 to 3 five- or six-memberedheterocyclic aryl groups selected from O, N and S, the substitutedsubstituents being selected from

and R₅; wherein ring A is preferably a 5 to 6-memberednitrogen-containing aromatic heterocyclic ring:

B is selected from —NH

wherein B₁ is from H, C₁₋₄ alkyl, halogenated C₁₋₄ alkyl, C₁₋₄ alkoxy,halogenated C₁₋₄ alkoxy;

R₁ is selected from a halogen atom, a C₁₋₆ alkyl group, a halogenatedC₁₋₆ alkyl group, a C₃₋₆ cycloalkyl group, a halogenated C₃₋₆ cycloalkylgroup, a C₁₋₆ alkoxy group, halogenated C₁₋₆ alkoxy, C₂₋₆ alkenyl, C₂₋₆hydroxy substituted alkenyl, C₂₋₆ alkynyl, C₂₋₆ hydroxy substitutedalkynyl, unsubstituted or substituted 5- or 6-membered aromatic oraromatic heterocyclic ring, said aromatic heterocyclic ring comprising 1to 3 hetero atoms selected from O, N, and S, the substitution being amono-, di- or tri-substitution, said substituent being selected from thegroup consisting of Ra; The aryl or heterocyclic aryl group ispreferably selected from the group consisting of a benzene ring, afuran, a thiophene, a pyrazole, a thiazole, and a pyrimidine;

Ra is selected from H, halogen, nitro, cyano, C₁₋₃ alkyl, halogenatedC₁₋₃ alkyl, —C(═O)ORb, —C(═O)NHRb, —NHRb, —ORb —NHCORb; Rb is selectedfrom H, C₁₋₃ alkyl, halogenated C₁₋₃ alkyl, C₁₋₃ alkoxy, halogenatedC₁₋₃ alkoxy, C₁₋₇ alkylamine;

R₂ is selected from the group consisting of H, —NHRc, —N(Rc)₂, —ORc,—SRc; Rc is selected from the group consisting of C₁₋₇ alkyl,halogenated C₁₋₇ alkyl, C₁₋₇ hydroxyalkyl, C₁₋₇ aminoalkyl;

R₃ is selected from the group consisting of halogen, nitro, cyano, C₁₋₃alkyl, halogenated C₁₋₃ alkyl, C₁₋₃ alkoxy, halogenated C₁₋₃ alkoxy,C₁₋₃ alkylamino, halogenated C₁₋₃ alkylamino group;

L₁ is selected from O, S, NH or a deletion;

m=0˜2;

R₄ is selected from C₁₋₇ alkyl, halogenated C₁₋₇ alkyl, hydroxysubstituted C₁₋₇ alkyl, C₁₋₇ alkylamino, halogenated C₁₋₇ alkylamino,C₁₋₇ alkoxy, halogenated C₁₋₇ alkoxy group, five- to eight-memberednitrogen-containing aliphatic heterocyclic ring;

R₅ is selected from the group consisting of halogen, nitro, cyano,trifluoromethyl, C₁₋₃ alkyl, C₁₋₃ alkoxy, amide and substituted alkylamide.

Preferably, the 2-polysubstituted aromatic ring-pyrimidine derivativecomprises a structure of formula II:

and an optical isomer thereof or a pharmaceutically acceptable salt orsolvate thereof, wherein: W, X, Y and Z are identical or different andare independently selected from N, C and O;

B is selected from —NH

wherein B₁ is selected from the group consisting of H, C₁₋₄ alkyl,halogenated C₁₋₄ alkyl, C₁₋₄ alkoxy, halogenated C₁₋₄ alkoxy;

R₁ is selected from halogen atom, C₁₋₆ alkyl group, halogenated C₁₋₆alkyl group, C₃₋₆ cycloalkyl group, halogenated C₃₋₆ cycloalkyl group,C₁₋₆ alkoxy group, halogenated C₁₋₆ alkoxy, C₂₋₆ alkenyl, C₂₋₆ hydroxysubstituted alkenyl, C₂₋₆ alkynyl, C₂₋₆ hydroxy substituted alkynyl,unsubstituted or substituted 5- or 6-membered aromatic or aromaticheterocyclic ring, said aromatic heterocyclic ring comprising 1 to 3hetero atoms selected from O, N and S, the substitution being a mono-,di- or tri-substitution, said substituent being selected from the groupconsisting of Ra;

Ra is selected from H, halogen, nitro, cyano, C₁₋₃ alkyl, halogenatedC₁₋₃ alkyl, —C(═O)ORb, —C(═O)NHRb, —NHRb, —ORb —NHCORb; Rb is selectedfrom H, C₁₋₃ alkyl, halogenated C₁₋₃ alkyl, C₁₋₃ alkoxy, halogenatedC₁₋₃ alkoxy, C₁₋₇ alkylamine;

R₂ is selected from the group consisting of H, —NHRc, —N(Rc)₂, —ORc,—SRc; Rc is selected from the group consisting of C₁₋₇ alkyl,halogenated C₁₋₇ alkyl, C₁₋₇ hydroxyalkyl, C₁₋₇ alkylamino group, C₁₋₇alkoxy group;

R₃ is selected from the group consisting of halogen, nitro, cyano, C₁₋₃alkyl, halogenated C₁₋₃ alkyl, C₁₋₃ alkoxy, halogenated C₁₋₃ alkoxy,C₁₋₃ alkylamino, halogenated C₁₋₃ alkylamino group;

L₁ is selected from O, S, NH or a deletion;

m=0˜2;

R₄ is selected from the group consisting of H, C₁₋₇ alkyl, halogenatedC₁₋₇ alkyl, hydroxy substituted C₁₋₇ alkyl, C₁₋₇ alkylamino, halogenatedC₁₋₇ alkylamino, C₁₋₇alkoxy group, halogenated C₁₋₇ alkoxy group, andfive- to eight-membered nitrogen-containing aliphatic heterocyclic ring;

R₅ is selected from the group consisting of halogen, nitro, cyano,trifluoromethyl, C₁₋₃ alkyl, C₁₋₃ alkoxy, amide, substituted alkylamide.

Preferably, the 2-polysubstituted aromatic ring-pyrimidine derivative,comprises a structure of formula III:

and an optical isomer thereof or a pharmaceutically acceptable salt orsolvate thereof;

wherein W, X, Y and Z are the same or different and are eachindependently selected from N or C;

B is selected from —NH,

wherein B₁ is selected from H, C₁₋₄ alkyl, halogenated C₁₋₄ alkyl, C₁₋₄alkoxy, halogenated C₁₋₄ alkoxy;

R₁ is selected from halogen atom, C₁₋₆ alkyl group, halogenated C₁₋₆alkyl group, C₃₋₆ cycloalkyl group, halogenated C₃₋₆ cycloalkyl group,C₁₋₆ alkoxy group, halogenated C₁₋₆ alkoxy, C₂₋₆ alkenyl, C₂₋₆ hydroxysubstituted alkenyl, C₂₋₆ alkynyl, C₂₋₆ hydroxy substituted alkynyl,unsubstituted or substituted 5- or 6-membered aromatic or aromaticheterocyclic ring, said aromatic heterocyclic ring comprising 1 to 3hetero atoms selected from O, N and S, the substitution being a mono-,di- or tri-substitution, said substituent being selected from the groupconsisting of Ra;

R_(a) is selected from H, halogen, nitro, cyano, C₁₋₃ alkyl, halogenatedC₁₋₃ alkyl, —C(═O)ORb, —C(═O)NHRb, —NHRb, —ORb —NHCORb; Rb is selectedfrom H, C₁₋₃ alkyl, halogenated C₁₋₃ alkyl, C₁₋₃ alkoxy, halogenatedC₁₋₃ alkoxy, C₁₋₇ alkylamine;

R₂ is selected from the group consisting of H, —NHRc, —N(Rc)₂, —ORc,—SRc; Rc is selected from the group consisting of C₁₋₇ alkyl,halogenated C₁₋₇ alkyl, C₁₋₇ hydroxyalkyl, C₁₋₇ alkylamine group, C₁₋₇alkoxy group;

R₃ is selected from the group consisting of halogen, nitro, cyano, C₁₋₃alkyl, halogenated C₁₋₃ alkyl, C₁₋₃ alkoxy, halogenated C₁₋₃ alkoxy,C₁₋₃ alkylamino, halogenated C₁₋₃ alkylamino group;

L₁ is selected from O, S, NH or a deletion;

m=0˜2;

R₄ is selected from the group consisting of H, C₁₋₇ alkyl, halogenatedC₁₋₇ alkyl, hydroxy substituted C₁₋₇ alkyl, C₁₋₇ alkylamino, halogenatedC₁₋₇ alkylamino, C₁₋₇ alkoxy group, halogenated C₁₋₇ alkoxy group, andfive- to eight-membered nitrogen-containing aliphatic heterocyclic ring;

R₅ is selected from the group consisting of halogen, nitro, cyano,trifluoromethyl, C₁₋₃ alkyl, C₁₋₃ alkoxy, amide, substituted alkylamide.

Preferably, the 2-polysubstituted aromatic ring-pyrimidine derivativecomprises a structure of formula IV:

and an optical isomer thereof or a pharmaceutically acceptable salt orsolvate thereof, wherein:

W, X, Y and Z are the same or different and are independently selectedfrom N, C and O;

R₁ is selected from halogen atom, C₁₋₆ alkyl group, halogenated C₁₋₆alkyl group, C₃₋₆ cycloalkyl group, halogenated C₃₋₆ cycloalkyl group,C₁₋₆ alkoxy group, halogenated C₁₋₆ alkoxy, C₂₋₆ alkenyl, C₂₋₆ hydroxysubstituted alkenyl, C₂₋₆ alkynyl, C₂₋₆ hydroxy substituted alkynyl,unsubstituted or substituted 5- or 6-membered aromatic or aromaticheterocyclic ring, said aromatic heterocyclic ring comprising 1 to 3hetero atoms selected from O, N and S, the substitution being a mono-,di- or tri-substitution, said substituent being selected from the groupconsisting of Ra;

R_(a) is selected from H, halogen, nitro, cyano, C₁₋₃ alkyl, halogenatedC₁₋₃ alkyl, —C(═O)ORb, —C(═O)NHRb, —NHRb, —ORb —NHCORb; Rb is selectedfrom H, C₁₋₃ alkyl, halogenated C₁₋₃ alkyl, C₁₋₃ alkoxy, halogenatedC₁₋₃ alkoxy, C₁₋₇ alkylamine;

R₂ is selected from the group consisting of H, —NHRc, —N(Rc)₂, —ORc,—SRc; Rc is selected from the group consisting of C₁₋₇ alkyl,halogenated C₁₋₇ alkyl, C₁₋₇ hydroxyalkyl, C₁₋₇ alkylamino group, C₁₋₇alkoxy group;

R₃ is selected from the group consisting of halogen, nitro, cyano, C₁₋₃alkyl, halogenated C₁₋₃ alkyl, C₁₋₃ alkoxy, halogenated C₁₋₃ alkoxy,C₁₋₃ alkylamino, halogenated C₁₋₃ alkylamino group;

L₁ is selected from O, S, NH or a deletion;

m=0-2;

R₄ is selected from the group consisting of H, C₁₋₇ alkyl, halogenatedC₁₋₇ alkyl, hydroxy substituted C₁₋₇ alkyl, C₁₋₇ alkylamino, halogenatedC₁₋₇ alkylamino, C₁₋₇ alkoxy group, halogenated C₁₋₇ alkoxy group, andfive- to eight-membered nitrogen-containing aliphatic heterocyclic ring;

R₅ is selected from the group consisting of halogen, nitro, cyano,trifluoromethyl, C₁₋₃ alkyl, C₁₋₃ alkoxy, amide, substituted alkylamide.

Preferably, in the 2-polysubstituted aromatic ring-pyrimidinederivative, the compound is selected from the group consisting of:

-   N⁴-methyl-5-(1-methyl-1H-pyrazol-4-yl)-N²-(1-methyl-5-(piperidin-4-yloxy)pyrazol-3-yl)-2,4-diaminopyrimidine;-   N⁴-methyl-5-(1-methyl-1H-pyrazol-4-yl)-N²-(3-(piperidin-4-yloxy)isoxazol-5-yl)-2,4-diaminopyrimidine;-   N⁴-methyl-5-(1-methyl-1H-pyrazol-4-yl)-N²-(1-methyl-2-(piperidin-4-yloxy)imidazol-4-yl)-2,4-diaminopyrimidine;-   N⁴-methyl-5-(1-methyl-1H-pyrazol-4-yl)-N²-(2-(piperidin-4-yloxy)isothiazol-5-yl)-2,4-diaminopyrimidine;-   N⁴-methyl-5-(1-methyl-1H-pyrazol-4-yl)-N²-(2-(piperidin-4-yloxy)-4,5-2H-oxazoline-5-yl)-2,4-diaminopyrimidine;    and-   N⁴-methyl-5-(1-methyl-1H-pyrazol-4-yl)-N²-(2-(piperidin-4-yloxy)thiazol-5-yl)-2,4-diaminopyrimidine.

Preferably, the 2-polysubstituted aromatic ring-pyrimidine derivative,comprises a structure of formula V:

and an optical isomer thereof or a pharmaceutically acceptable salt orsolvate thereof, wherein:

W, X, Y and Z are the same or different and are each independentlyselected from N or C;

R₁ is selected from halogen atom, C₁₋₆ alkyl group, halogenated C₁₋₆alkyl group, C₃₋₆ cycloalkyl group, halogenated C₃₋₆ cycloalkyl group,C₁₋₆ alkoxy group, halogenated C₁₋₆ alkoxy, C₂₋₆ alkenyl, C₂₋₆ hydroxysubstituted alkenyl, C₂₋₆ alkynyl, C₂₋₆ hydroxy substituted alkynyl,unsubstituted or substituted 5- or 6-membered aromatic or aromaticheterocyclic ring, said aromatic heterocyclic ring comprising 1 to 3hetero atoms selected from O, N and S, the substitution being a mono-,di- or tri-substitution, said substituent being selected from the groupconsisting of Ra;

Ra is selected from H, halogen, nitro, cyano, C₁₋₃ alkyl, halogenatedC₁₋₃ alkyl, —C(═O)ORb, —C(═O)NHRb, —NHRb, —ORb —NHCORb; Rb is selectedfrom H, C₁₋₃ alkyl, halogenated C₁₋₃ alkyl, C₁₋₃ alkoxy, halogenatedC₁₋₃ alkoxy, C₁₋₇ alkylamine;

R₂ is selected from the group consisting of H, —NHRc, —N(Rc)₂, —ORc,—SRc; Rc is selected from the group consisting of C₁₋₇ alkyl,halogenated C₁₋₇ alkyl, C₁₋₇ hydroxyalkyl, C₁₋₇ alkane Amino group, C₁₋₇alkoxy group;

R₃ is selected from the group consisting of halogen, nitro, cyano, C₁₋₃alkyl, halogenated C₁₋₃ alkyl, C₁₋₃ alkoxy, halogenated C₁₋₃ alkoxy,C₁₋₃ alkylamino, halogenated C₁₋₃ alkylamino group;

L₁ is selected from O, S, NH or a deletion;

m=0˜2;

R₄ is selected from the group consisting of H, C₁₋₇ alkyl, halogenatedC₁₋₇ alkyl, hydroxy substituted C₁₋₇ alkyl, C₁₋₇ alkylamino, halogenatedC₁₋₇ alkylamino, C₁₋₇ alkoxy group, halogenated C₁₋₇ alkoxy group, andfive- to eight-membered nitrogen-containing aliphatic heterocyclic ring;

R₅ is selected from the group consisting of halogen, nitro, cyano,trifluoromethyl, C₁₋₃ alkyl, C₁₋₃ alkoxy, amide, substituted alkylamide.

In the 2-polysubstituted aromatic ring-pyrimidine derivative, thecompound is selected from the group consisting of:

-   5-phenyl-N-(2-cyano-3-(piperidin-3-oxy)pyridin-5-yl)-2-aminopyrimidine;-   5-(3-fluorophenyl)-N-(2-cyano-3-(piperidin-3-yloxy)pyridin-5-yl)-2-aminopyrimidine;-   5-(4-fluorophenyl)-N-(2-cyano-3-(piperidin-3-yloxy)pyridin-5-yl)-2-aminopyrimidine;-   5-(3-methoxyphenyl)-N-(2-cyano-3-(piperidin-3-yloxy)pyridin-5-yl)-2-aminopyrimidine;-   5-(4-methoxyphenyl)-N-(2-cyano-3-(piperidin-3-yloxy)pyridin-5-yl)-2-aminopyrimidine;-   5-(pyridin-3-yl)-N-(2-cyano-3-(piperidin-3-oxy)pyridin-5-yl)-2-aminopyrimidine;-   5-(pyridin-4-yl)-N-(2-cyano-3-(piperidin-3-yloxy)pyridin-5-yl)-2-aminopyrimidine;-   5-(thien-2-yl)-N-(2-cyano-3-(piperidin-3-yloxy)pyridin-5-yl)-2-aminopyrimidine;-   5-(furan-2-yl)-N-(2-cyano-3-(piperidin-3-yloxy)pyridin-5-yl)-2-aminopyrimidine;-   5-trifluoromethyl-N-(2-cyano-3-(piperidin-3-oxy)pyridin-5-yl)-2-aminopyrimidine;-   (R)-5-(1-methyl-1H-pyrazol-4-yl)-N-(2-cyano-3-(piperidin-3-yloxy)pyridin-5-yl)-2-aminopyrimidine;-   (S)-5-(1-methyl-1H-pyrazol-4-yl)-N-(2-cyano-3-(piperidin-3-yloxy)pyridin-5-yl)-2-aminopyrimidine;-   4-methoxy-5-(3-fluorophenyl)-N-(2-cyano-3-(piperidin-4-methyl)oxypyridin-5-yl)-2-aminopyrimidine;-   4-methoxy-5-(1-methyl-1H-pyrazol-4-yl)-N-(2-cyano-3-(piperidin-4-methyl)oxypyridin-5-yl)2-aminopyrimidine;-   4-methoxy-5-(3-fluorophenyl)-N-(2-cyano-3-(2-dimethylaminoethoxy)pyridin-5-yl)-2-aminopyrimidine;-   4-methoxy-5-(1-methyl-1H-pyrazol-4-yl)-N-(2-cyano-3-(2-dimethylaminoethoxy)pyridine-5-yl)    2-aminopyrimidine;-   4-methoxy-5-trifluoromethyl-N-(2-cyano-3-(piperidin-4-methyl)oxypyridin-5-yl)-2-aminopyrimidine;-   N⁴-methyl-5-phenyl-N²-(2-cyano-3-(piperidin-3-oxy)pyridin-5-yl)-2,4-diaminopyrimidine;-   N⁴-methyl-5-(3-fluorophenyl)-N²-(2-cyano-3-(piperidin-3-yloxy)pyridin-5-yl)-2,4-diaminopyrimidine;-   N⁴-methyl-5-(4-fluorophenyl)-N²-(2-cyano-3-(piperidin-3-yloxy)pyridin-5-yl)-2,4-diaminopyrimidine;-   N⁴-methyl-5-(2-fluorophenyl)-N²-(2-cyano-3-(piperidin-3-oxy)pyridin-5-yl)-2,4-diaminopyrimidine;-   N⁴-methyl-5-(3-methoxyphenyl)-N²-(2-cyano-3-(piperidin-3-yloxy)pyridin-5-yl)-2,4-diaminopyrimidine;-   N⁴-methyl-5-(4-methoxyphenyl)-N²-(2-cyano-3-(piperidin-3-yloxy)pyridin-5-yl)-2,4-diaminopyrimidine;-   N⁴-methyl-5-(2,4-dimethoxyphenyl)-N²-(2-cyano-3-(piperidin-3-yloxy)pyridin-5-yl)-2,4-diaminopyrimidine;-   N⁴-methyl-5-(pyridin-3-yl)-N²-(2-cyano-3-(piperidin-3-yloxy)pyridin-5-yl)-2,4-diaminopyrimidine;-   N⁴-methyl-5-(pyridin-4-yl)-N²-(2-cyano-3-(piperidin-3-yloxy)pyridin-5-yl)-2,4-diaminopyrimidine;-   N⁴-methyl-5-(thiophen-2-yl)-N²-(2-cyano-3-(piperidin-3-oxy)pyridin-5-yl)-2,4-diaminopyrimidine;-   N⁴-methyl-5-(furan-2-yl)-N²-(2-cyano-3-(piperidin-3-yloxy)pyridin-5-yl)-2,4-diaminopyrimidine;-   N⁴-methyl-5-(5-chloro-furan-2-yl)-N²-(2-cyano-3-(piperidin-3-yloxy)pyridin-5-yl)-2,4-diaminopyrimidine;-   N₄-methyl-5-(5-methoxycarbonylthiophen-2-yl)-N²-(2-cyano-3-(piperidin-3-yloxy)pyridin-5-yl)-2,4-diaminopyrimidine;-   N⁴-methyl-5-(1-methyl-1H-pyrazol-4-yl)-N²-(2-cyano-3-(piperidin-3-yloxy)pyridin-5-yl)-2,4-diaminopyrimidine;-   N⁴-methyl-5-(1-methyl-1H-pyrazol-5-yl)-N²-(2-cyano-3-(piperidin-3-yloxy)pyridin-5-yl)-2,4-diaminopyrimidine;-   N⁴-methyl-5-trifluoromethyl-N²-(2-cyano-3-(piperidin-3-oxy)pyridin-5-yl)-2,4-diaminopyrimidine;-   N⁴-methyl-5-(1-methyl-1H-pyrazol-4-yl)-N²-(2-cyano-3-(piperidin-4-methyl)oxypyridin-5-yl)-2,4-diaminopyrimidine;-   (R)—N⁴-methyl-5-(1-methyl-1H-pyrazol-4-yl)-N²-(2-cyano-3-(piperidin-3-yloxy)pyridine-5-yl)-2,4-diaminopyrimidine;-   (S)—N⁴-methyl-5-(1-methyl-1H-pyrazol-4-yl)-N²-(2-cyano-3-(piperidin-3-yloxy)pyridine-5-yl)-2,4-diaminopyrimidine;-   (R)—N⁴-methyl-5-trifluoromethyl-N²-(2-cyano-3-(piperidin-3-oxy)pyridin-5-yl)-2,4-diaminopyrimidine;-   (S)—N⁴-methyl-5-trifluoromethyl-N²-(2-cyano-3-(piperidin-3-oxy)pyridin-5-yl)-2,4-diaminopyrimidine;-   (R)—N⁴-methyl-5-(1-methyl-1H-pyrazol-4-yl)-N²-(2-cyano-3-(pyrrole-3-oxy)pyridin-5-yl)-2,4-diaminopyrimidine;-   (S)—N⁴-methyl-5-(1-methyl-1H-pyrazol-4-yl)-N²-(2-cyano-3-(pyrrole-3-oxy)pyridin-5-yl)-2,4-diaminopyrimidine;-   N⁴-methyl-5-(1-methyl-1H-pyrazol-4-yl)-N²-(2-cyano-3-(piperidin-4-yloxy)pyridin-5-yl)-2,4-diaminopyrimidine;-   N⁴-methyl-5-(1-methyl-1H-pyrazol-4-yl)-N²-(2-cyano-3-(2-dimethylaminoethoxy)pyridin-5-yl)-2,4-diaminopyrimidine;-   (R)—N⁴-methyl-5-(1-methyl-1H-pyrazol-4-yl)-N²-(2-cyano-3-(1-dimethylaminopropyl-2-oxy)    pyridin-5-yl)-2,4-diaminopyrimidine;-   (S)—N⁴-methyl-5-(1-methyl-1H-pyrazol-4-yl)-N²-(2-cyano-3-(1-dimethylaminopropyl-2-oxy)    pyridin-5-yl)-2,4-diaminopyrimidine;-   N⁴-methyl-5-(1-methyl-1H-pyrazol-4-yl)-N²-(2-cyano-3-(N-methylpiperidin-4-yloxy)pyridine-5-yl)2,4-diaminopyrimidine;-   N⁴-methyl-5-trifluoromethyl-N²-(2-cyano-3-(N-methylpiperidin-4-yloxy)pyridin-5-yl)-2,4-diaminopyrimidine;-   N⁴-methyl-5-(4-methylthiazol-2-yl)-N²-(2-cyano-3-(piperidin-3-yloxy)pyridin-5-yl)-2,4-diaminopyrimidine;

and a pharmaceutically acceptable salt or solvate thereof.

The invention adopts a method familiar to technicians in this field toprepare salt of 2-pyrimidine derivatives described in the invention. Thesalt may be an organic acid salt, a mineral acid salt and so on, and theorganic acid salt includes a citrate, a fumarate, an oxalate, a malate,a L-malate, and a D-malate, lactate, camphorsulfonate,p-toluenesulfonate, methanesulfonate, benzoate, etc.; the inorganic acidsalt includes a hydrohalide, a sulfate, a phosphate, a nitrate, and soon. For example, with a lower alkylsulfonic acid such as methanesulfonicacid, trifluoromethanesulfonic acid or the like may form a mesylatesalt, a triflate salt; and an arylsulfonic acid such as benzenesulfonicacid or p-toluenesulfonic acid; with organic carboxylic acids such asacetic acid, fumaric acid, tartrate, L-tartaric acid, D-tartaric acid,oxalic acid, maleic acid, malate, L-malic acid, D-malic acid, succinicacid or citric acid and so on may form corresponding salts; and with anamino acid such as glutamic acid or aspartic acid may form a glutamateor an aspartate. Corresponding salts may also be formed with inorganicacids such as hydrohalic acids (e.g., hydrofluoric acid, hydrobromicacid, hydroiodic acid, hydrochloric acid), nitric acid, carbonic acid,sulfuric acid or phosphoric acid.

The second purpose of the invention is to provide a pharmaceuticalcomposition comprising at least one active ingredient together with oneor more pharmaceutically acceptable carriers or excipients, said activeingredient it may be a 2-substituted pyrimidine compound of the presentinvention, an optical isomer of the compound, a solvate of the compoundor an optical isomer thereof in a pharmaceutically acceptable salt, thecompound or an optical isomer thereof Any one or any of a variety ofthem.

The carrier includes conventional diluents, excipients, fillers,binders, wetting agents, disintegrating agents, absorption enhancers,surfactants, adsorption carriers, lubricants, etc. in the pharmaceuticalfield, etc. and fragrances, sweetener may also be added if necessary.The medicament of the present invention can be prepared into variousforms such as tablets, powders, granules, capsules, oral liquids andinjectable preparations, and the medicaments of the above respectivedosage forms can be prepared according to a conventional method in thepharmaceutical field.

The present invention also provides a compound of the formula (I) to theformula (V), and an optical isomer thereof, or a pharmaceuticallyacceptable salt or solvate thereof, and an optical isomer thereof, or apharmaceutically acceptable compound thereof. The use of accepted saltsor solvates, alone and/or in combination with radiation therapy, otherdrugs, in the preparation of Chk1 inhibitors, particularly in thepreparation of therapeutically proliferative diseases. The cellproliferative diseases include tumors, and the tumors are breast cancer,ovarian cancer, narcoma, lung cancer, prostate cancer, colon cancer,rectal cancer, renal cancer, pancreatic cancer, blood cancer, lymphoma,neuroblastoma, and glioma, head cancer, neck cancer, thyroid cancer,liver cancer, vulvar cancer, cervical cancer, endometrial cancer,testicular cancer, bladder cancer, esophageal cancer, stomach cancer,nasopharyngeal cancer, buccal cancer, oral cancer, gastrointestinalstromal tumor, skin cancer, multiple myeloma. The antitumor agent whichcan be used in combination with the compound provided by the presentinvention or a pharmaceutically acceptable salt intended includes, butis not limited to, at least one of the following classes: antimetabolite(gemcitabine, 5-fluorouracil, hydroxyurea, pemetrexed);bioalkylatingagengts (eg cisplatin, carboplatin); topoisomeraseinhibitors (irinotecan, doxorubicin); small molecule inhibitors (MEKinhibitors, PARP inhibitors, Scr kinase inhibitors, mTOR inhibitors,farnesyltransferase inhibitors, etc.).

Another object of the present invention is to provide a process for thepreparation of the above target compound by the following steps:

First Method:

(1) 5-Bromo-2-substituted-3-nitropyridine (or5-bromo-2-cyano-3-nitropyridine) was reacted with different fattyalcohols under basic conditions (in the presence of NaH) to obtaindifferent substituted pyridine fragments;

(2) Starting with 5-bromo-2,4-dichloropyrimidine, followed by methyletherification or methylamination to give 5-bromo-2-chloro-4-substitutedpyrimidine then was ammoniated to get5-bromo-N⁴-methylpyrimidine-2,4-diamine or5-bromo-4-methoxypyrimidin-2-amine. 2-Aminopyrimidine was brominated toobtain 5-bromo-2-aminopyrimidine. The Suzuki-Miyaura coupling reactionwas occurred between 5-bromo-N⁴-methylpyrimidine-2,4-diamine or5-bromo-4-methoxypyrimidin-2-amine or 5-bromo-2-aminopyrimidine andcorresponding borate or boric acid and then Buchwald-Hartwigcross-coupling was occurred between the product of the Suzuki-Miyauracoupling reaction and the substituted pyridine fragment from the firststep, followed by acidic deprotection to obtain the target compound; or5-bromo-N⁴-methylpyrimidine-2,4-diamine was firstly carried outBuchwald-Hartwig cross-coupling with the substituted pyridine fragmentfrom the first step, then followed by Suzuki-Miyaura coupling reactionwith corresponding borate or boric acid, and then deprotection underacidic conditions to obtain the target compound; Compound 1-6 wasprepared according to the following synthetic route:

Compound 7-16 was prepared according to the following synthetic route:

Compound 17-43 was prepared according to the following synthetic route:

Compound 44 was prepared according to the following synthetic route:

Second Method:

(1) 5-Bromo-2-cyano-3-nitropyridine was reacted with different fattyalcohols under basic conditions (conditions of NaH) to obtain differentsubstituted pyridine fragments;

(2) 2,4-Dichloro-5-trifluoromethylpyrimidine as starting material,followed by methyl etherification or methylamination, then ammoniationto obtain 2,4-disubstituted-5-trifluoromethylpyrimidine.2-Chloro-5-trifluoromethylpyrimidine was ammoniated to get2-amino-5-trifluoromethylpyrimidine. Buchwal-Hartwig cross-coupling wascarried out between 2,4-disubstituted-5-trifluoromethylpyrimidine or2-amino-5-trifluoromethylpyrimidine with the substituted pyridinefragment from the first step, and finally removing the Boc protectinggroup to obtain the target compound.

The synthetic route of compound 46-50 is as follows:

The inventors of the present invention have experimentally confirmedthat most of the compounds of the present invention have moderate tostrong Chk1 kinase inhibitory activity and are useful for the treatmentof solid tumors or blood cancers associated with cell proliferation inhumans or animals. A series of novel small molecule Chk1 inhibitors weredesigned and synthesized by using 2-aminopyrimidine as a lead compoundobtained through structure-based virtual screening, and Chk1 kinaseinhibitory activity of these compounds was tested. The results showedthat most of the compounds exhibited moderate to strong Chk1 inhibitoryactivity and were promising Chk1 inhibitors, providing new drugs forcancer therapy.

BRIEF DESCRIPTION OF THE DRAWINGS

The FIGURE is a diagram showing activity of CHK1 inhibitor combiningwith other drugs on MV 4-11 cell line.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENT

Further description of the present invention was illustrated combiningwith the preferred embodiments. The embodiments below are exemplary onlyand not intended to be limiting.

Embodiment of Preparation 1N⁴-methyl-5-(1-methyl-1H-pyrazol-4-yl)-N²-(1-methyl-5-(piperidin-4-yloxy)-pyrazol-3-yl)-2,4-diaminopyrimidine (Compound 1)

Step 1: synthesis ofN-tert-butoxycarbonyl-4-((3-bromo-1-methyl-1H-pyrazol-5-yl) oxy)piperidin (Intermediate 1-2)

N-tert-Butyloxycarbonyl-4-hydroxypiperidine (460 mg, 2.29 mmol) wasdissolved into anhydrous THF (7.2 mL), cooling in ice bath, adding 60%sodium hydride (108 mg, 4.5 mmol) portionwise and stirring at roomtemperature for 10 min; The mixture was heated to 35° C. and stirred for10 min; Subsequently, the above sodium solution was added dropwiseslowly (completed in about 20 min) to the solution of3-bromo-1-methyl-5-nitropyrazole (360 mg, 1.75 mmol) in anhydrous THF4.8 mL in a three-necked flask under protection of nitrogen and stirredat room temperature for 1 h. The reaction was quenched by the additionof a saturated ammonium chloride solution, and the solvent wasevaporated under reduced pressure. The resultant product was purified bysilica gel column chromatography eluting PE:EtOAc (50:9) to give ayellow liquid. Yield: 79%; LCMS: m/z=361 [M+1]⁺.

Step 2, Synthesis of 4-methylamino-2-chloro-5-bromopyrimidine(Intermediate 1-4)

To a solution of 5-bromo-2,4-dichloropyrimidine (5 g, 22 mmol) inmethanol (42 mL), 33% methylamine alcohol solution (6.75 mL) was addeddropwise under an ice water bath, and the reaction mixture was stirredat room temperature 30 min. The solvent was evaporated under reducedpressure and the crude product was purified by column chromatography(eluent PE:EtOAc=5:1) to give a white solid. Yield: 88%; mp: 139-141°C.; ¹H NMR (500 MHz, DMSO-d₆): δ 8.85 (s, Ar—H, 1H), 7.75 (br, NH, 1H),2.85 (d, J=3.9 Hz, CH₃, 3H); ESI-MS: m/z=222 [M+1]⁺.

Step 3. Synthesis of 5-bromo-N⁴-methyl-2,4-diaminopyrimidine(Intermediate 1-5)

Compound 1-4 (275 mg, 1.23 mmol) was placed in a sealed tube, and asolution of ammonia-saturated ethanol (20 mL) was added and the reactionmixture was stirred at 100° C. for 24 h. After cooling to roomtemperature, the solvent was removed by evaporation. The residue waspurified by silica gel column chromatography (eluent PE:EtOAc=2:1) togive a white solid. Yield: 78%; ¹H NMR (500 MHz, CDCl³): δ 7.86 (s,Ar—H, 1H), 5.22 (br, NH, 1H), 4.85 (br, NH₂, 2H), 2.99 (d, J=6.0 Hz,CH₃, 3H); ESI-MS: m/z=204 [M+1]⁺.

Step 4. Synthesis of5-(1-methyl-1H-pyrazol-4-yl)-N⁴-methyl-2,4-diaminopyrimidine(Intermediate 1-6)

Compound 1-5 (290 mg, 1.43 mmol), 1-methyl-1H-pyrazole-4-boronic acidpinacol ester (358 mg, 1.72 mmol) and Pd(dppf)Cl₂ (54 mg, 0.07 mmol)were placed in reaction flask. Then ethylene glycol dimethyl ether (14mL) and IN aqueous Na₂CO₃ solution were added. The reaction was carriedout under refluxing overnight. After cooling, the solvent was removedunder reduced pressure. The residue was purified by columnchromatography on silica gel (eluent CH₂Cl₂:EtOH=25:1) to give a whitesolid. Yield: 75%; mp: 226-228° C.; ¹H NMR (500 MHz, DMSO-d₆): δ 7.88(s, Ar—H, 1H), 7.49 (s, Ar—H, 1H), 7.35 (s, Ar—H, 1H), 6.22 (br, NH₂,2H), 5.96 (q, J=4.5 Hz, NH, 1H), 3.51 (s, CH₃, 3H), 2.86 (d, J=4.5 Hz,CH₃, 3H); ESI-MS: m/z=205 [M+1]⁺.

Step 5. Synthesis ofN⁴-methyl-5-(1-methyl-1H-pyrazol-4-yl)-N²-(1-methyl-5-(piperidin-4-yloxy)pyrazol-3-yl)-2,4-diaminopyrimidine(Compound 1)

A mixture of compound 1-6 (169 mg, 0.986 mmol), compound 1-2 (376 mg,0.986 mmol), tris(dibenzylideneacetone)dipalladium (9 mg, 0.00986 mmol),4,5-bisdiphenylphosphine-9,9-dimethyloxanium (15 mg, 0.026 mmol, Cs₂CO₃(450 mg, 1.38 mmol) in dry dioxane (6 mL) was heated to reflux overnightunder nitrogen. The cooled reaction mixture was filtered and thefiltrate was removed by evaporation. The obtained residue was purifiedby column chromatography on silica gel (eluent CH₂Cl₂:EtOH=30:1) to givea white solid. Removal of the Boc protecting group with trifluoroaceticacid gave a white solid. Yield: 65%; LCMS: m/z=385 [M+1]⁺.

Preparative Example 2N⁴-Methyl-5-(1-methyl-1H-pyrazol-4-yl)-N²-(3-(piperidin-4-yloxy)isoxazol-5-yl)-2,4-diaminopyrimidine(Compound 2)

Step 1. Synthesis ofN-tert-butoxycarbonyl-4-((5-bromo-isoxazol-3-yl)oxy)piperidine(Intermediate 1-8)

Synthetic Procedure Reference Example 1, Step 1. Compounds 1-8 wereprepared from (Compound 1-7) according to the same procedure as Compound1-2. Yield: 63%; LCMS: m/z=348 [M+1]⁺.

Step 2.N⁴-Methyl-5-(1-methyl-1H-pyrazol-4-yl)-N²-(3-(piperidin-4-yloxy)isoxazol-5-yl)-2,4-diaminopyrimidine(Compound 2)

The synthetic procedure was carried out according to the same procedureas in PREPARATIVE EXAMPLE 1, Step 5. Compound 2 was prepared by asynthetic method similar to Compound 1, using Intermediates 1-8 and 1-6as starting materials. Yield: 65%; LCMS: m/z=371 [M+1]⁺.

Preparative Example 3.N⁴-Methyl-5-(1-methyl-1H-pyrazol-4-yl)-N²-(1-methyl-2-(piperidin-4-yloxy)imidazol-4-yl)-2,4-diaminopyrimidine(Compound 3)

Step 1. Synthesis ofN-tert-butoxycarbonyl-4-((4-bromo-1-methyl-1H-imidazol-2-yl)oxy)piperidine(Intermediate 1-10)

Synthetic Procedure Reference Example 1, Step 1. Compound 1-10 wasprepared from 4-bromo-1-methyl-2-nitroimidazole (Compound 1-9) accordingto the same procedure as Compound 1-2. Yield: 67%; LCMS: m/z=361 [M+1]⁺.

Step 2.N⁴-Methyl-5-(1-methyl-1H-pyrazol-4-yl)-N²-(1-methyl-2-(piperidin-3-yloxy)imidazol-4-yl)-2,4-diaminopyrimidine(Compound 3)

The synthetic procedure was carried out according to the same procedureas in PREPARATIVE EXAMPLE 1, Step 5. Compound 3 was prepared by asynthetic method similar to Compound 1, using Intermediates 1-10 and 1-6as starting materials. Yield: 65%; LCMS: m/z=384 [M+1]⁺.

Preparative Example 4.N⁴-Methyl-5-(1-methyl-1H-pyrazol-4-yl)-N²-(2-(piperidin-4-yloxy)isothiazol-5-yl)-2,4-diaminopyrimidine(Compound 4)

Step 1. Synthesis ofN-tert-butoxycarbonyl-4-((5-bromo-isothiazol-3-yl)oxy)piperidine(Intermediate 1-12)

Synthetic procedure Reference to Example 1, Step 1. Compound 1-12 wasprepared from 5-bromo-3-nitroisothiazole (Compound 1-11) according tothe same procedure as Compound 1-2. Yield: 66%; LCMS: m/z=364[M+1]⁺.

Step 2.N⁴-Methyl-5-(1-methyl-1H-pyrazol-4-yl)-N²-(2-(piperidin-4-yloxy)isothiazol-5-yl)-2,4-diaminopyrimidine (Compound 4)

The synthetic procedure was carried out according to the same procedureas in PREPARATIVE EXAMPLE 1, Step 5. Compound 4 was prepared by asynthetic method similar to Compound 1, using Intermediates 1-12 and 1-6as starting materials. Yield: 65%; LCMS: m/z=387 [M+1]⁺.

Preparative Example 5.N⁴-Methyl-5-(1-methyl-1H-pyrazol-4-yl)-N²-(2-(piperidin-4-yloxy)-4,5-(2H-oxazolin-5-yl)-2,4-diaminopyrimidine(Compound 5)

Step 1. Synthesis ofN-tert-butoxycarbonyl-4-((5-bromo-4,5-2H-oxazolin-2-yl)oxy)piperidine(Intermediate 1-14)

Synthetic Procedure Reference Example 1, Step 1. Compound 1-14 wasprepared from 5-bromo-2-nitro-4,5-2H-oxazoline (compound 1-13) accordingto the same procedure as compound 1-2. Yield: 67%; LCMS: m/z=361 [M+1]⁺.

Step 2.N⁴-Methyl-5-(1-methyl-1H-pyrazol-4-yl)-N²-(2-(piperidin-4-yloxy)-4,5-2H-oxazolin-5-yl)-2,4-diaminopyrimidine(Compound 5)

The synthetic procedure was carried out according to the same procedureas in PREPARATIVE EXAMPLE 1, Step 5. Compound 5 was prepared by asynthetic method similar to Compound 1, using Intermediates 1-14 and 1-6as starting materials. Yield: 65%; LCMS: m/z=373 [M+1]⁺.

Preparative Example 6.N⁴-Methyl-5-(1-methyl-1H-pyrazol-4-yl)-N²-(2-(piperidin-4-yloxy)thiazol-5-yl)-2,4-diaminopyrimidine(Compound 6)

Step 1. Synthesis ofN-tert-butoxycarbonyl-4-((5-bromo-thiazol-2-yl)oxy)piperidine(Intermediate 1-16)

Synthetic Procedure Reference Example 1, Step 1. Compound 1-16 wasprepared from 5-bromo-2-nitrothiazole (Compound 1-15) according to thesame procedure as Compound 1-2. Yield: 63%; LCMS: m/z=364 [M+1]⁺.

Step 2.N⁴-Methyl-5-(1-methyl-1H-thiazol4-yl)-N²-(1-methyl-2-(piperidin-4-yloxy)pyrazol-4-yl)-2,4-diaminopyrimidine(Compound 6)

The synthetic procedure was carried out according to the same procedureas in PREPARATIVE EXAMPLE 1, Step 5. Compound 6 was prepared by asynthetic method similar to Compound 1, using Intermediates 1-16 and 1-6as starting materials. Yield: 65%; LCMS: m/z=387 [M+1]⁺.

Preparative Example 7.5-Phenyl-N-(2-cyano-3-(piperidin-3-oxy)pyridin-5-yl)-2-aminopyrimidine(Compound 7)

Step 1. Synthesis of2-cyano-5-bromo-3-(N-tert-butoxycarbonylpiperidin-3-yloxy)pyridine(Intermediate 2-2)

Synthetic Procedure Reference to Example 1, Step 1. Compound 2-2 wasprepared according to the same procedure as Compound 1-2, using5-bromo-3-nitro-2-cyanopyridine (Compound 2-1) andN-tert-butoxycarbonyl-3-hydroxypiperidine as starting materials. Yield:79%; ¹H NMR (500 MHz, DMSO-d₆): δ 8.26 (s, Ar—H, 1H), 7.53 (s, Ar—H,1H), 4.38 (br, CH, 1H), 3.63 (br, CH, 1H), 3.51 (br, CH, 1H), 3.38 (br,CH, 2H), 1.98-1.96 (m, CH, 1H), 1.91-1.85 (m, CH, 2H), 1.51 (br, CH,1H), 1.36 (s, CH₃×3, 9H); ESI-MS: m/z=382 [M+1]⁺.

Step 2. Synthesis of 5-bromo-2-aminopyrimidine (Intermediate 2-4)

2-Aminopyrimidine (2.5 g, 26.29 mmol) was dissolved in acetonitrile (25mL). Then N-bromosuccinimide (4.6 g, 27.9 mmol) was added in ice-bath.The reaction mixture was stirred at room temperature overnight. Themixture was evaporated under reduced pressure, and the resultant productwas washed with water (100 mL) and filtered with suction. The obtainedproduct was dried in vacuum to get a white solid. Yield: 97%; mp:241-243° C.

Step 3. Synthesis of 2-amino-5-phenylpyrimidine (Intermediate 2-5)

Synthetic Procedure Reference Example 1, Step 4. Compound 2-5 wasprepared by using intermediate 2-4 and phenylboronic acid as startingmaterials. Yield: 86%; mp: 159-161° C.; ¹H NMR (400 MHz, DMSO-d₆): δ8.57 (s, Ar—H, 2H), 7.62 (d, J=7.6 Hz, Ar—H, 2H), 7.45 (t, J=7.2 Hz,Ar—H, 2H), 7.34 (t, J=7.2 Hz, Ar—H, 1H), 6.77 (s, NH₂, 2H); ESI-MS:m/z=172 [M+1]⁺.

Step 4. Synthesis of5-phenyl-N-(2-cyano-3-(piperidin-3-oxy)pyridin-5-yl)-2-aminopyrimidine(Compound 7)

Synthetic Procedure Reference Example 1, Step 5. Compound 7 was preparedusing Intermediates 2-5 and 2-2 as starting materials. Yield: 65%; mp:84-86° C.; ¹H NMR (500 MHz, DMSO-d₆): δ 10.55 (s, NH, 1H), 8.99 (s,Ar—H, 2H), 8.65 (d, J=2.0 Hz, Ar—H, 1H), 8.39 (d, J=2.0 Hz, Ar—H, 1H),7.78 (d, J=7.0 Hz, Ar—H, 2H), 7.52 (t, J=7.5 Hz, Ar—H, 2H), 7.43 (t,j=7.0 Hz, Ar—H, 1H), 4.42-4.37 (m, CH, 1H), 3.19 (d, j=12.0 Hz, 2.0 Hz,CH₂, 1H), 2.81-2.77 (m, CH₂, 1H), 2.66-2.62 (m, CH₂, 1H), 2.56-2.53 (m,CH₂, 1H), 2.14-2.11 (m, CH₂, 1H), 1.76-1.71 (m, CH₂, 1H), 1.65-1.58 (m,CH₂, 1H), 1.54-1.46 (m, CH₂, 1H); ¹³C NMR (100 MHZ, DMSO-d₆): δ 160.22,158.43, 157.73, 153.05, 142.82, 137.60, 133.83, 129.37, 116.42, 113.77,111.72, 106.92, 104.53, 86.99, 57.06, 45.54, 38.63, 28.17; ESI-MS:m/z=373 [M+1]⁺.

Preparative Example 8.5-(3-Fluorophenyl)-N-(2-cyano-3-(piperidin-3-yloxy)pyridin-5-yl)-2-aminopyrimidine(Compound 8)

Step 1. Synthesis of 2-amino-5-(3-fluorophenyl)pyrimidine (Intermediate2-6)

Synthetic Procedure Reference Example 1, Step 4. Compound 2-6 wasprepared from Intermediate 2-4 and 3-fluorophenylboronic acid to give awhite solid. Yield: 83%; mp: 168-170° C.; ¹H NMR (400 MHz, DMSO-d₆): δ8.62 (s, Ar—H, 2H), 7.53-7.43 (m, Ar—H, 3H), 7.16-7.11 (m, Ar—H, 1H),6.86 (s, NH₂, 2H); ESI-MS: m/z=190 [M+1]⁺.

Step 2. Synthesis of5-(3-fluorophenyl)-N-(2-cyano-3-(piperidin-3-yloxy)pyridin-5-yl)-2-aminopyrimidine(Compound 8)

Synthetic Procedure Reference Example 1, Step 5. Compound 8 was preparedfrom Intermediates 2-6 and 2-2 to give a white solid. Yield: 72%; mp:107-109° C.; NMR (500 MHz, DMSO-d₆): δ 10.59 (br, NH, 1H), 9.03 (s,Ar—H, 2H), 8.65 (d, J=1.5 Hz, Ar—H, 1H), 8.37 (s, Ar—H, 1H), 7.70 (d,J=10.5 Hz, Ar—H, 1H), 7.65 (d, J=8.0 Hz, Ar—H, 1H), 7.56 (dd, J=14.0 Hz,8.0 Hz, Ar—H, 1H), 7.26 (td, J=8.5 Hz, 2.5 Hz, Ar—H, 1H), 4.42-4.37 (m,CH, 1H), 3.19 (dd, J=12.0 Hz, 2.0 Hz, CH₂, 1H), 2.82-2.78 (m, CH₂, 1H),2.67-2.63 (m, CH₂, 1H), 2.57-2.52 (m, CH₂, 1H), 2.14-2.11 (m, CH₂, 1H),1.77-1.71 (m, CH₂, 1H), 1.66-1.59 (m, CH₂, 1H), 1.54-1.47 (m, CH₂, 1H);ESI-MS: m/z=391 [M+1]⁺.

Preparative Example 9.5-(4-Fluorophenyl)-N-(2-cyano-3-(piperidin-3-oxy)pyridin-5-yl)-2-aminopyrimidine(Compound 9)

Step 1. Synthesis of 2-amino-5-(4-fluorophenyl)pyrimidine (Intermediate2-7)

Synthetic Procedure Reference Example 1, Step 4. Compound 2-7 wasprepared from the intermediate 2-4 and 4-fluorophenylboronic acid togive a white solid. Yield: 85%; mp: 172-174° C.; ¹H NMR (400 MHz,DMSO-d₆): δ 8.58 (s, Ar—H, 2H), 7.68-7.64 (m, Ar—H, 2H), 7.29 (t, J=8.8Hz, Ar—H, 2H), 6.78 (s, NH₂, 2H); ESI-MS: m/z=190 [M+1]⁺.

Step 2. Synthesis of4-(4-fluorophenyl)-N-(2-cyano-3-(piperidin-3-yloxy)pyridin-5-yl)-2-aminopyrimidine(Compound 9)

Synthetic Procedure Reference Example 1, Step 5. Compound 9 was preparedfrom intermediates 2-7 and 2-2 to give a white solid. Yield: 78%; mp:182-184° C.; ¹H NMR (500 MHz, DMSO-d₆): δ 10.54 (br, NH, 1H), 8.96 (s,Ar—H, 2H), 8.65 (d, J=2.0 Hz, Ar—H, 1H), 8.36 (s, Ar—H, 1H), 7.83 (dd,J=8.5 Hz, 5.5 Hz, Ar—H, 2H), 7.35 (t, J=8.0 Hz, Ar—H, 2H), 4.41-4.36 (m,CH, 1H), 3.20 (dd, j=12.0 Hz, 2.5 Hz, CH₂, 1H), 2.82-2.78 (m, CH₂, 1H),2.67-2.63 (m, CH₂, 1H), 2.57-2.52 (m, CH₂, 1H), 2.14-2.11 (m, CH₂, 1H),1.77-1.71 (m, CH₂, 1H), 1.65-1.58 (m, CH₂, 1H), 1.54-1.46 (m, CH₂, 1H);¹³C NMR (100 MHz, DMSO-d₆): δ 163.09, 161.14, 158.27, 157.25, 155.90,141.82, 134.15, 130.59, 128.17, 128.11, 125.15, 116.21, 116.08, 115.91,113.84, 109.21, 74.69, 49.65, 45.28, 29.82, 24.27; ESI-MS: m/z=391[M+1]⁺.

Preparative Example 10.5-(3-Methoxyphenyl)-N-(2-cyano-3-(piperidin-3-oxy)pyridin-5-yl)-2-aminopyrimidine(Compound 10)

Step 1. Synthesis of 2-amino-5-(3-methoxyphenyl)pyrimidine (Intermediate2-8)

Synthetic Procedure Reference Example 1, Step 4. Compound 2-8 wasprepared from intermediate 2-4 and 3-methoxyphenylboronic acid to give awhite solid. Yield: 87%; mp: 133-135° C.; ¹H NMR (400 MHz, DMSO-d₆): δ8.58 (s, Ar—H, 2H), 7.36 (t, J=8.0 Hz, Ar—H, 1H), 7.18 (d, J=7.6 Hz,Ar—H, 2H), 6.90 (dd, J=8.4 Hz, 1.6 Hz, Ar—H, 1H), 6.78 (s, NH₂, 2H),3.81 (s, CH₃, 3H); ESI-MS: m/z=202 [M+1]⁺.

Step 2. Synthesis of5-(3-methoxyphenyl)-N-(2-cyano-3-(piperidin-3-oxy)pyridin-5-yl)-2-aminopyrimidine(Compound 10)

Synthetic Procedure Reference Example 1, Step 5. Compound 10 wasprepared from intermediates 2-8 and 2-2 to give a white solid. Yield:74%; mp: 102-104° C.; ¹H NMR (500 MHz, DMSO-d₆): δ 10.54 (br, NH, 1H),8.98 (s, Ar—H, 2H), 8.65 (d, J=2.0 Hz, Ar—H, 1H), 8.37 (d, J=2.0 Hz,Ar—H, 1H), 7.42 (t, J=8.0 Hz, Ar—H, 1H), 7.33-7.32 (m, Ar—H, 2H),6.99-6.97 (m, Ar—H, 1H), 4.42-4.37 (m, CH, 1H), 3.85 (s, CH₃, 3H), 3.20(dd, J=12.0 Hz, 2.0 Hz, CH₂, 1H), 2.82-2.78 (m, CH₂, 1H), 2.68-2.64 (m,CH₂, 1H), 2.57-2.53 (m, CH₂, 1H), 2.14-2.11 (m, CH₂, 1H), 1.77-1.71 (m,CH₂, 1H), 1.66-1.59 (m, CH₂, 1H), 1.55-1.47 (m, CH₂, 1H); ¹³C NMR (100MHz, DMSO-d₆): δ 159.92, 158.41, 157.25, 156.03, 141.83, 135.43, 134.18,130.23, 125.86, 118.19, 116.23, 113.84, 113.65, 111.37, 109.21, 74.66,55.22, 49.63, 45.29, 29.81, 24.24; ESI-MS: m/z=403 [M+1]⁺.

Preparative Example 115-(4-Methoxyphenyl)-N-(2-cyano-3-(piperidin-3-oxy)pyridin-5-yl)-2-aminopyrimidine(Compound 11)

Step 1. Synthesis of 2-amino-5-(4-methoxyphenyl)pyrimidine (Intermediate2-9)

Synthetic Procedure Reference Example 1, Step 4. Compound 2-9 wasprepared from intermediate 2-4 and 4-methoxyphenylboronic acid to give awhite solid. Yield: 82%; mp: 165-167° C.; ¹H NMR (400 MHz, DMSO-d₆): δ8.51 (s, Ar—H, 2H), 7.55 (d, J=8.8 Hz, Ar—H, 2H), 7.01 (d, J=8.8 Hz,Ar—H, 2H), 6.67 (s, NH₂, 2H), 3.78 (s, CH₃, 3H); ESI-MS: m/z=202 [M+1]⁺.

Step 2. Synthesis of5-(4-methoxyphenyl)-N-(2-cyano-3-(piperidin-3-yloxy)pyridin-5-yl)-2-aminopyrimidine(Compound 11)

Synthetic Procedure Reference Example 1, Step 5. Compound 11 wasprepared from intermediates 2-9 and 2-2 to give a white solid. Yield:75%; mp: 218-220° C.; ¹H NMR (400 MHz, DMSO-d₆): δ 10.47 (s, NH, 1H),8.91 (s, Ar—H, 2H), 8.64 (s, Ar—H, 1H), 8.36 (s, Ar—H, 1H), 7.71 (d,J=8.8 Hz, Ar—H, 2H), 7.06 (d, J=8.8 Hz, Ar—H, 2H), 4.40-4.36 (m, CH,1H), 3.81 (s, CH₃, 3H), 3.20 (d, J=12.0 Hz, CH₂, 1H), 2.81-2.78 (m, CH₂,1H), 2.67-2.62 (m, CH₂, 1H), 2.56-2.51 (m, CH₂, 1H), 2.14-2.12 (m, CH₂,1H), 1.76-1.72 (m, CH₂, 1H), 1.66-1.57 (m, CH₂, 1H), 1.54-1.46 (m, CH₂,1H); ¹³C NMR (100 MHz, DMSO-d₆): δ 159.25, 157.86, 157.26, 155.34,141.92, 134.08, 127.20, 126.29, 125.85, 116.24, 114.61, 113.65, 109.00,74.69, 55.20, 49.69, 45.31, 29.84, 24.31; ESI-MS: m/z=403 [M+1]⁺.

Preparative Example 125-(Pyridin-3-yl)-N-(2-cyano-3-(piperidin-3-oxy)pyridin-5-yl)-2-aminopyrimidine(Compound 12)

Step 1. Synthesis of 2-amino-5-(pyridin-3-yl)pyrimidine (Intermediate2-10)

Synthetic Procedure Reference Example 1, Step 4. Compound 2-10 wasprepared from intermediate 2-4 and pyridine-3-boronic acid to give awhite solid. Yield: 79%; mp: 183-185° C.; ¹H NMR (400 MHz, DMSO-d₆): δ8.87 (s, Ar—H, 1H), 8.64 (s, Ar—H, 2H), 8.53 (br, Ar—H, 1H), 8.05 (d,J=7.6 Hz, Ar—H, 1H), 7.46 (d, J=4.0 Hz, Ar—H, 1H), 6.89 (s, NH₂, 2H);ESI-MS: m/z=173 [M+1]⁺.

Step 2. Synthesis of5-(pyridin-3-yl)-N-(2-cyano-3-(piperidin-3-yloxy)pyridin-5-yl)-2-aminopyrimidine(Compound 12)

Synthetic Procedure Reference Example 1, Step 5. Compound 12 wasprepared from intermediates 2-10 and 2-2 to give a white solid. Yield:75%; mp: 221-223° C.; ¹H NMR (500 MHz, DMSO-d₆): δ 10.61 (br, NH, 1H),9.06 (s, Ar—H, 2H), 9.00 (d, J=2.5 Hz, Ar—H, 1H), 8.67 (d, J=1.5 Hz,Ar—H, 1H), 8.62 (dd, J=4.5 Hz, 1.5 Hz, Ar—H, 1H), 8.37 (d, J=2.0 Hz,Ar—H, 1H), 8.21 (td, J=8.0 Hz, 2.0 Hz, Ar—H, 1H), 7.54 (dd, J=8.0 Hz,4.0 Hz, Ar—H, 1H), 4.43-4.39 (m, CH, 1H), 3.20 (dd, J=12.0 Hz, 2.0 Hz,CH₂, 1H), 2.83-2.78 (m, CH₂, 1H), 2.69-2.65 (m, CH₂, 1H), 2.59-2.54 (m,CH₂, 1H), 2.15-2.12 (m, CH₂, 1H), 1.78-1.72 (m, CH₂, 1H), 1.67-1.60 (m,CH₂, 1H), 1.55-1.47 (m, CH₂, 1H); ESI-MS: m/z=374 [M+1]⁺.

Preparative Example 135-(Thien-2-yl)-N-(2-cyano-3-(piperidin-3-oxy)pyridin-5-yl)-2-aminopyrimidine(Compound 13)

Step 1. Synthesis of 2-amino-5-(thien-2-yl)pyrimidine (Intermediate2-11)

Synthetic Procedure Reference Example 1, Step 4. Compound 2-11 wasprepared from intermediate 2-4 and thienyl-2-boronic acid to give awhite solid. Yield: 84%; mp: 156-158° C.; ¹H NMR (400 MHz, DMSO-d₆): δ8.53 (s, Ar—H, 2H), 7.49 (d, J=4.8 Hz, Ar—H, 1H), 7.38 (d, J=2.8 Hz,Ar—H, 1H), 7.12 (t, J=4.4 Hz, Ar—H, 1H), 6.87 (s, NH₂, 2H): ESI-MS:m/z=178 [M+1]⁺.

Step 2. Synthesis of5-(thien-2-yl)-N-(2-cyano-3-(piperidin-3-yloxy)pyridin-5-yl)-2-aminopyrimidine(Compound 13)

Synthetic Procedure Reference Example 1, Step 5. Compound 13 wasprepared from intermediates 2-11 and 2-2 to give a white solid. Yield:74%; mp: 212-214° C.; ¹H NMR (500 MHz, DMSO-d₆): δ 10.58 (br, NH, 1H),8.93 (s, Ar—H, 2H), 8.61 (d, J=2.0 Hz, Ar—H, 1H), 8.35 (d, J=2.0 Hz,Ar—H, 1H), 7.63-7.61 (m, Ar—H, 2H), 7.20 (dd, J=5.0 Hz, 3.5 Hz, Ar—H,1H), 4.42-4.37 (m, CH, 1H), 3.19 (dd, J=12.0 Hz, 2.0 Hz, CH₂, 1H),2.82-2.78 (m, CH₂, 1H), 2.67-2.63 (m, CH₂, 1H), 2.57-2.52 (m, CH₂, 1H),2.14-2.10 (m, CH₂, 1H), 1.77-1.71 (m, CH₂, 1H), 1.66-1.59 (m, CH₂, 1H),1.54-1.46 (m, CH₂, 1H); ¹³C NMR (100 MHz, DMSO-d₆): δ 158.00, 157.25,154.63, 141.67, 136.46, 134.18, 128.55, 126.15, 124.27, 121.13, 116.20,113.91, 109.24, 74.64, 49.63, 45.30, 29.81, 24.26; ESI-MS: m/z=379[M+1]⁺.

Preparative Example 145-(Furan-2-yl)-N-(2-cyano-3-(piperidin-3-oxy)pyridin-5-yl)-2-aminopyrimidine(Compound 14)

Step 1. Synthesis of 2-amino-5-(furan-2-yl)pyrimidine (Intermediate2-12)

Synthetic Procedure Reference Example 1, Step 4. Compound 2-12 wasprepared from intermediate 2-4 and furan-2-boronic acid to give a whitesolid. Yield: 83%; mp: 156-158° C.; ¹H NMR (400 MHz, DMSO-d₆): δ 8.57(s, Ar—H, 2H), 7.69 (s, Ar—H, 1H), 6.88 (s, NH, 2H), 6.78 (d, J=2.0 Hz,Ar—H, 1H), 6.56 (s, Ar—H, 1H); ESI-MS: m/z=162 [M+1]⁺

Step 2. Synthesis of5-(furan-2-yl)-N-(2-cyano-3-(piperidin-3-yloxy)pyridin-5-yl)-2-aminopyrimidine(Compound 14)

Synthetic Procedure Reference Example 1, Step 5. Compound 14 wasprepared from intermediates 2-12 and 2-2 to give a white solid. Yield:70%; mp: 200-202° C.; ¹H NMR (400 MHz, DMSO-d₆): δ 10.58 (br, NH, 1H),8.95 (s, Ar—H, 2H), 8.59 (d, J=1.2 Hz, Ar—H, 1H), 8.36 (s, Ar—H, 1H),7.81 (s, Ar—H, 1H), 7.05 (d, J=2.8 Hz, Ar—H, 1H), 6.65 (t, J=1.6 Hz,Ar—H, 1H), 4.41-4.37 (m, CH, 1H), 3.20 (d, J=12 Hz, CH₂, 1H), 2.82-2.79(m, CH₂, 1H), 2.67-2.62 (m, CH₂, 1H), 2.57-2.55 (m, CH₂, 1H), 2.14-2.12(m, CH₂, 1H), 1.76-1.73 (m, CH₂, 1H), 1.67-1.58 (m, CH₂, 1H), 1.54-1.49(m, CH₂, 1H); ¹³C NMR (100 MHZ, DMSO-d₆): δ 157.76, 157.23, 153.18,148.23, 143.35, 141.62, 134.17, 117.90, 116.19, 113.89, 112.08, 109.23,106.19, 74.66, 49.63, 45.30, 29.81, 24.27; ESI-MS: m/z=363 [M+1]⁺

Preparative Example 15(R)-5-(1-Methyl-1H-pyrazol-4-yl)-N-(2-cyano-3-(piperidin-3-yloxy)pyridine-5-yl)-2-aminopyrimidine(Compound 15)

Step 1. Synthesis of(R)-2-cyano-5-bromo-3-(N-tert-butoxycarbonylpiperidin-3-yloxy)pyridine(Intermediate 2-13)

Synthetic Procedure Reference Example 1, Step 1. Compound 2-13 wasprepared from 5-bromo-3-nitro-2-cyanopyridine (Compound 2-1) and(R)—N-tert-butyloxycarbonyl-3-hydroxypiperidine as starting materialsaccording to the same procedure as Compound 1-2, and a yellow oil.Yield: 76%; ¹H NMR (500 MHz, DMSO-d₆): δ 8.26 (s, Ar—H, 1H), 7.53 (s,Ar—H, 1H), 4.38 (br, CH, 1H), 3.63 (br, CH, 1H), 3.51 (br, CH, 1H), 3.38(br, CH, 2H), 1.98-1.96 (m, CH, 1H), 1.91-1.85 (m, CH, 2H), 1.51 (br,CH, 1H), 1.36 (s, CH₃×3, 9H); ESI-MS: m/z=382 [M+1]⁺.

Step 2. Synthesis of 2-amino-5-(1-methyl-1H-pyrazol-4-yl)pyrimidine(Intermediate 2-14)

Synthetic Procedure Reference Example 1, Step 4. Compound 2-14 wasprepared from intermediate 2-4 and 1-methylpyrazol-4-boronic acidpinacol ester as a white solid. Yield: 85%; mp: 174-176° C.; ¹H NMR (400MHz, DMSO-d₆): δ 8.46 (s, Ar—H, 2H), 8.03 (s, Ar—H, 1H), 7.78 (s, Ar—H,1H), 6.57 (s, NH₂, 2H), 3.84 (s, CH₃, 3H); ESI-MS: m/z=176 [M+1]⁺.

Step 3. Synthesis of(R)-5-(1-methyl-1H-pyrazol-4-yl)-N-(2-cyano-3-(piperidin-3-yloxy)pyridin-5-yl)-2-aminopyrimidine(Compound 15)

Synthetic Procedure Reference Example 1, Step 5. Compound 15 wasprepared from intermediates 2-14 and 2-13 to give a white solid. Yield:73%; mp: 230° C. (decomposition); ¹H NMR (500 MHz, DMSO-d₆): δ 10.42(br, NH, 1H), 8.87 (s, Ar—H, 2H), 8.59 (d, J=1.5 Hz, Ar—H, 1H), 8.35 (d,J=2.0 Hz, Ar—H, 1H), 8.23 (s, Ar—H, 1H), 7.97 (s, Ar—H, 1H), 4.41-4.36(m, CH, 1H), 3.89 (s, CH₃, 3H), 3.19 (dd, J=12.0 Hz, 2.0 Hz, CH₂, 1H),2.82-2.78 (m, CH₂, 1H), 2.68-2.64 (m, CH₂, 1H), 2.58-2.53 (m, CH₂, 1H),2.14-2.11 (m, CH₂, 1H), 1.78-1.72 (m, CH₂, 1H), 1.65-1.58 (m, CH₂, 1H),1.54-1.46 (m, CH₂, 1H); ¹³C NMR (125 MHz, DMSO-d₆): δ 157.28, 154.26,141.98, 135.75, 134.02, 127.58, 119.77, 116.28, 115.46, 113.46, 108.76,74.59, 49.62, 45.27, 38.74, 29.78, 24.19; ESI-MS: m/z=377 [M+1]⁺.

Preparative Example 16(S)-5-(1-Methyl-1H-pyrazol-4-yl)-N-(2-cyano-3-(piperidin-3-yloxy)pyridine-5-yl-2-aminopyrimidine(Compound 16)

Step 1. Synthesis of(S)-2-cyano-5-bromo-3-(N-tert-butoxycarbonylpiperidin-3-yloxy)pyridine(Intermediate 2-15)

Synthetic Procedure Reference Example 1, Step 1. Compound 2-15 wasprepared from 5-bromo-3-nitro-2-cyanopyridine (Compound 2-1) and(S)—N-tert-butoxycarbonyl-3-hydroxypiperidine as starting materialsaccording to the same procedure as Compound 1-2, a yellow oil. Yield:78%; ¹H NMR (500 MHz, DMSO-d₆): δ 8.26 (s, Ar—H, 1H), 7.53 (s, Ar—H,1H), 4.38 (br, CH, 1H), 3.63 (br, CH, 1H), 3.51 (br, CH, 1H), 3.38 (br,CH, 2H), 1.98-1.96 (m, CH, 1H), 1.91-1.85 (m, CH, 2H), 1.51 (br, CH,1H), 1.36 (s, CH₃λ3, 9H); ESI-MS: m/z=382 [M+1]⁺.

Step 2. Synthesis of(N)-5-(1-methyl-1H-pyrazol-4-yl)-N-(2-cyano-3-(piperidin-3-yloxy)pyridin-5-yl)2-aminopyrimidine (Compound 16)

Synthetic Procedure Reference Example 1, Step 5. Compound 16 wasprepared from intermediates 2-14 and 2-15 to give a white solid. Yield:72%; mp: 230° C. (decomposition); ¹H NMR (500 MHz, DMSO-d₆): δ 10.42(br, NH, 1H), 8.87 (s, Ar—H, 2H), 8.59 (d, J=1.5 Hz, Ar—H, 1H), 8.35 (d,J=2.0 Hz, Ar—H, 1H), 8.23 (s, Ar—H, 1H), 7.97 (s, Ar—H, 1H), 4.41-4.36(m, CH, 1H), 3.89 (s, CH₃, 3H), 3.19 (dd, J=12.0 Hz, 2.0 Hz, CH₂, 1H),2.82-2.78 (m, CH₂, 1H), 2.68-2.64 (m, CH₂, 1H), 2.58-2.53 (m, CH₂, 1H),2.14-2.11 (m, CH₂, 1H), 1.78-1.72 (m, CH₂, 1H), 1.65-1.58 (m, CH₂, 1H),1.54-1.46 (m, CH₂, 1H); ¹³C NMR (125 MHz, DMSO-d₆): δ 157.28, 154.26,141.98, 135.75, 134.02, 127.58, 119.77, 116.28, 115.46, 113.46, 108.76,74.59, 49.62, 45.27, 38.74, 29.78, 24.19; ESI-MS: m/z=377 [M+1]⁺.

Preparative Example 174-Methoxy-5-(3-fluorophenyl)-N-(2-cyano-3-(piperidin-4-methyl)oxypyridin-5-yl)-2-aminopyrimidine(Compound 17)

Step 1. Synthesis of2-cyano-5-bromo-3-(N-tert-butoxycarbonylpiperidin-4-methyl)oxypyridine(Intermediate 2-16)

Synthetic Procedure Reference Example 1, Step 1. Compound 2-16 wasprepared from 5-bromo-3-nitro-2-cyanopyridine (Compound 2-1) andN-tert-butoxycarbonylpiperidin-4-methanol as starting materialsaccording to the same procedure as Compound 1-2. Yield: 80%; ¹H NMR (500MHz, CDCl₃): δ 8.34 (s, Ar—H, 1H), 7.51 (s, Ar—H, 1H), 4.20 (d, J=11.5Hz, CH₂, 2H), 3.95 (br, CH₂, 2H), 2.80 (br, CH₂, 2H), 2.14-2.05 (m, CH,1H), 1.89 (br, CH₂, 2H), 1.47 (s, CH₃×3, 9H), 1.33-1.26 (m, CH₂, 2H);ESI-MS: m/z=206 [M+1]⁺.

Step 2. Synthesis of 4-methoxy-2-chloro-5-bromopyrimidine (Intermediate2-17)

5-Bromo-2,4-dichloropyrimidine (500 mg, 2.194 mmol) was dissolved inanhydrous methanol (5 mL), and sodium methoxide (sodium 56 mg, 2.42mmol) in methanol (1.85 mL) was added to the above solution undernitrogen. The mixture was stirred at room temperature overnight. Thereaction was quenched by the addition of saturated ammonium chloridesolution and was evaporated at reduced pressure to remove the solvent.Then CH₂Cl₂ (30 mL) was added and was washed with water (30 mL). Theorganic layer was separated and evaporated under a reduced pressure toremove the solvent. The crude product was purified by silica gel columnchromatography (eluent PE:EtOAc=4:1) to give a white solid. Yield: 90%;¹H NMR (400 MHz, CDCl₃): δ 8.44 (s, Ar—H, 1H), 4.11 (s, CH₃, 3H);ESI-MS: m/z=223 [M+1]⁺.

Step 3. Synthesis of 4-methoxy-5-bromo-2-aminopyrimidine (Intermediate2-18)

Synthetic Procedure Reference Example 1, Step 3. Compound 2-18 wasprepared from intermediate 2-17 as a starting material according to thesame procedure as Compound 1-5, a white solid. Yield: 75%; ¹H NMR (500MHz, CDCl₃): δ 8.13 (s, Ar—H, 1H), 5.41 (s, NH₂, 2H), 3.91 (s, CH₃, 3H);ESI-MS: m/z=205 [M+1]⁺.

Step 4. Synthesis of 4-methoxy-5-(3-fluorophenyl)-2-aminopyrimidine(Intermediate 2-19)

Synthetic Procedure Reference Example 1, Step 4. Compound 2-19, a whitesolid, was prepared from intermediate 2-18 and 3-fluorophenylboronicacid as starting materials. Yield: 88%; mp: 127-128° C.; ¹H NMR (500MHz, CDCl₃): δ 7.98 (s, Ar—H, 1H), 7.46-7.42 (m, Ar—H, 1H), 7.18 (d,J=7.5 Hz, Ar—H, 1H), 7.12-7.08 (m, Ar—H, 2H), 5.19 (s, NH₂, 2H), 4.10(S, NH₂, 2H), 3.96 (S, CH₃, 3H); ESI-MS: m/z=220 [M+1]⁺.

Step 5. Synthesis of4-methoxy-5-(3-fluorophenyl)-N-(2-cyano-3-(piperidin-4-methyl)oxypyridin-5-yl)-2-aminopyrimidine(Compound 17)

Synthetic Procedure Reference Example 1, Step 5. Compound 17, a whitesolid, was prepared from intermediates 2-16 and 2-19. Yield: 79%; mp:139-141° C.; ¹H NMR (500 MHz, CDCl₃): δ 8.36 (d, J=2.0 Hz, Ar—H, 1H),8.18 (s, Ar—H, 1H), 8.08 (d, J=2.0 Hz, Ar—H, 1H), 7.58-7.54 (m, Ar—H,1H), 7.24-7.21 (m, Ar—H, 2H), 7.15-7.12 (m, Ar—H, 1H), 7.04 (br, NH,1H), 4.05 (s, CH₃, 3H), 3.97 (d, J=6.5 Hz, CH₂, 2H), 3.20-3.18 (m, CH₂,2H), 2.73-2.72 (m, CH₂, 2H), 2.13-2.06 (m, CH, 1H), 1.93-1.91 (m, CH₂,2H), 1.40-1.32 (m, CH₂, 2H); ¹³C NMR (125 MHz, DMSO-d₆): δ 163.85,163.46, 161.52, 158.92, 158.03, 157.93, 141.22, 136.25, 135.99, 135.92,131.07, 131.01, 125.40, 116.22, 116.05, 115.84, 114.98, 114.82, 114.64,114.14, 111.54, 72.92, 55.50, 44.11, 34.25, 27.19; ESI-MS: m/z=435[M+1]⁺.

Preparative Example 184-Methoxy-5-(3-fluorophenyl)-N-(2-cyano-3-(piperidin-4-methyl)oxypyridin-5-yl)-2-aminopyrimidine(Compound 18)

Step 1. Synthesis of4-methoxy-5-(1-methyl-1H-pyrazol-4-yl)-2-aminopyrimidine (Intermediate2-20)

Synthetic Procedure Reference Example 1, Step 4. Compound 2-20, a whitesolid, was prepared from intermediate 2-18 and 1-methylpyrazol-4-boronicacid pinacol ester. Yield: 85%; ¹H NMR (500 MHz, CDCl₃): δ 7.96 (s,Ar—H, 1H), 7.58 (s, Ar—H, 1H), 7.47 (s, Ar—H, 1H), 5.18 (s, NH₂, 2H),3.97 (s, CH₃, 3H), 3.94 (s, CH₃, 3H); ESI-MS: m/z=206 [M+1]⁺.

Step 2. Synthesis of4-methoxy-5-(1-methyl-1H-pyrazol-4-yl)-N-(2-cyano-3-(piperidin-4-methyl)oxypyridin-5-yl)-2-aminopyrimidine(Compound 18)

Synthetic Procedure Reference Example 1, Step 5. Compound 18, a whitesolid, was prepared from intermediates 2-16 and 2-20. Yield: 82%; mp:213-215° C.; ¹H NMR (500 MHz, DMSO-d₆): δ 9.02 (s, Ar—H, 1H), 8.69 (d,J=2.0 Hz, Ar—H, 1H), 8.25 (s, Ar—H, 1H), 8.24 (d, j=2.0 Hz, Ar—H, 1H),8.05 (s, NH, 1H), 7.70 (s, Ar—H, 1H), 5.33 (s, NH, 1H), 4.05 (d, j=6.5Hz, CH₂, 2H), 3.92 (s, CH₃, 3H), 3.91 (s, CH₃, 3H), 3.31-3.28 (m, CH₂,2H), 2.93-2.87 (m, CH₂, 2H), 2.19-2.11 (m, CH, 1H), 1.93-1.91 (m, CH₂,2H), 1.55-1.47 (m, CH₂, 2H); ESI-MS: m/z=421 [M+1]⁺.

Preparative Example 19

Preparation of Compounds 19 to 31.

Step 1. Synthesis of Intermediates 2-21-2-33

The synthetic procedure was described as Example 1, Step 4. Compounds2-21-2-33 were prepared from the intermediate 1-5 and the correspondingboronic acid to obtain white solid.

Step 2. Synthesis of Compound 19-31

The synthetic procedure was described as Example 1, Step 5. Compounds19-31 were prepared from the intermediates 2-21 to 2-33 and 2-2 to givewhite solids.

The synthetic method of compound 30 is as follows:

Synthetic Procedure Reference Example 1, Step 5. Intermediates forcompound was prepared from intermediates 2-31 and 2-2, and the Bocprotected intermediate (100 mg, 0.203 mmol) was dissolved in DMF/THF(1:1) mixed solution (2 mL), N-chlorosuccinimide (27 mg, 0.203 mmol) wasadded drop wise, and stirred at 60° C. for 5 hours. The solvent wasevaporated under reduced pressure. AcOEt was added, washed with water.The organic layer was combined, dried over anhydrous sodium sulfate andevaporated under reduced pressure to get resultant product, then the Bocprotected group was removed with trifluoroacetic acid to afford yellowsolid.

Preparative Example 20.N⁴-Methyl-5-(1-methyl-1H-pyrazol-4-yl)-N²-(2-cyano-3-(piperidin-3-yloxy)pyridin-5-yl)-2,4-diaminopyrimidine(Compound 32)

Step 1. Synthesis ofN⁴-methyl-5-(1-methyl-1H-pyrazol-4-yl)-N²-(2-cyano-3-(piperidin-3-yloxy)pyridin-5-yl-2,4-diaminopyrimidine(Compound 32)

Synthetic Procedure Reference Example 1, Step 5. Compound 32 wasprepared from intermediates 1-6 and 2-2 to give a white solid. Yield:79%; mp: 207-209° C.; ¹H NMR (500 MHz, DMSO-d₆): δ 9.92 (s, NH, 1H),8.51 (s, Ar—H, 2H), 7.91 (s, Ar—H, 1H), 7.90 (s, Ar—H, 1H), 7.61 (s,Ar—H, 1H), 6.74 (q, J=4.5 Hz, NH, 1H), 4.40-4.35 (m, CH, 1H), 3.89 (s,CH₃, 3H), 3.15 (dd, J=12.0 Hz, 2.0 Hz, CH₂, 1H), 2.94 (d, J=5.0 Hz, CH₃,3H), 2.79-2.74 (m, CH₂, 1H), 2.63-2.59 (m, CH₂, 1H), 2.54-2.48 (m, CH₂,1H), 2.11-2.07 (m, CH₂, 1H), 1.74-1.68 (m, CH₂, 1H), 1.62-1.55 (m, CH₂,1H), 1.45-1.37 (m, CH₂, 1H); ESI-MS: m/z=406 [M+1]⁺.

Preparative Example 21.N⁴-Methyl-5-(1-methyl-1H-pyrazol-5-yl)-N²-(2-cyano-3-(piperidin-3-yloxy)pyridin-5-yl)-2,4-diaminopyrimidine(Compound 33)

Step 1. Synthesis of5-(1-methyl-1H-pyrazol-5-yl)-N⁴-methyl-2,4-diaminopyrimidine(Intermediate 2-35)

Synthetic Procedure Reference Example 1, Step 4. Compound 2-35 wasprepared from intermediate 1-5 and 1-methyl-1H-pyrazole-5-boronic acidpinacol ester as a white solid. Yield: 65%; ¹H NMR (500 MHz, DMSO-d₆): δ7.54 (s, Ar—H, 1H), 7.47 (d, J=1.5 Hz, Ar—H, 1H), 6.24 (br, NH₂, 2H),6.20 (d, J=1.5 Hz, Ar—H, 1H), 6.07 (q, J=4.5 Hz, NH, 1H), 3.61 (s, CH₃,3H), 2.75 (d, J=4.5 Hz, CH₃, 3H); ESI-MS: m/z=205 [M+1]⁺.

Step 2. Synthesis ofN⁴-methyl-5-(1-methyl-1H-pyrazol-4-yl)-N²-(2-cyano-3-(piperidin-3-yloxy)pyridin-5-yl-2,4-diaminopyrimidine(Compound 33)

Synthetic Procedure Reference Example 1, Step 5. Compound 33 wasprepared from intermediates 2-35 and 2-2 to give a white solid. Yield:74%; ¹H NMR (400 MHz, DMSO-d₆): δ 10.07 (br, NH, 1H), 8.55 (s, Ar—H,1H), 8.51 (s, Ar—H, 1H), 7.87 (s, Ar—H, 1H), 7.54 (d, J=1.6 Hz, Ar—H,1H), 6.78 (q, J=4.0 Hz, NH, 1H), 6.32 (d, J=1.6 Hz, Ar—H, 1H), 4.41-4.36(m, CH, 1H), 3.67 (s, CH₃, 3H), 3.16 (dd, j=9.6 Hz, 2.0 Hz, CH₂, 1H),2.93 (d, j=3.6 Hz, CH₃, 3H), 2.79-2.75 (m, CH₂, 1H), 2.64-2.59 (m, CH₂,1H), 2.54-2.49 (m, CH₂, 1H), 2.11-2.07 (m, CH₂, 1H), 1.74-1.68 (m, CH₂,1H), 1.63-1.56 (m, CH₂, 1H), 1.45-1.37 (m, CH₂, 1H); ¹³C NMR (100 MHz,DMSO-d₆): δ 160.55, 159.15, 157.30, 155.63, 142.41, 138.23, 135.67,134.32, 116.34, 113.22, 108.60, 107.38, 101.53, 74.30, 49.78, 45.27,36.50, 29.89, 28.14, 24.20; ESI-MS: m/z=406 [M+1]⁺.

Preparative Example 22. Preparation of Compounds 34 to 43

Step 1. Synthesis of Intermediates 2-36˜2-42

Synthetic Procedure Reference Example 1, Step 1. Intermediates 2-36-2-42were prepared by using 5-bromo-3-nitro-2-cyanopyridine (compound 2-1)and the corresponding alcohol as starting materials through a syntheticmethod similar to compound 1-2.

Step 2. Synthesis of Compound 34-43

Synthetic Procedure Reference Example 1, Step 5. Compounds 34-43 wereprepared from 2-13, 2-15, 2-16, 2-36˜2-42 and intermediate 2-34,respectively, to give white solids.

Preparative Example 23.N⁴-Methyl-5-(4-methylthiazol-2-yl)-N²-(2-cyano-3-(piperidin-3-yloxy)pyridin-5-yl)-2,4-diaminopyrimidine(Compound 44)

Step 1. Synthesis of lithium trisisopropyl(4-methylthiazol-2-yl)borate(intermediate 2-44)

4-Methylthiazole (1 g, 10.09 mmol), triisopropyl borate (2.35 mL, 10.09mmol) was dissolved in a mixture of anhydrous toluene and THF (32 mL,v/v, 4:1) under nitrogen. Cooled to −78° C., n-butyl lithium (3.83 mL,2.5 mol/L, 9.58 mmol) was added slowly within 85 min, and the reactionwas stirred for 135 min. Slowly warmed to 0° C. (about 1.5 h),isopropanol (2.84 mL) was added and stirred overnight. The solvent wasevaporated under reduced pressure, and anhydrous acetone (17 mL) wasadded, then evaporated. The mixture was filtered under a nitrogenatmosphere, washed with acetonitrile of 55° C., dried over vacuum toobtain a white solid, which was to be used directly in the next stepwithout further purification.

Step 2. Synthesis ofN⁴-methyl-5-bromo-N²-(2-cyano-3-(N-tert-butoxycarbonylpiperidin-3-yloxy)pyridin-5-yl)-2,4-diaminopyrimidine(intermediate 2-45)

The synthetic procedure was described as Example 1, Step 5. Theintermediate 2-45 was prepared from intermediates 1-5 and 2-2 to give awhite solid. Yield: 70%; NMR (400 MHz, DMSO-d₆): δ 10.05 (s, NH, 1H),8.46 (s, Ar—H, 1H), 8.41 (s, Ar—H, 1H), 8.13 (s, Ar—H, 1H), 7.32 (q,J=4.5 Hz, NH, 1H), 4.59 (br, CH, 1H), 3.96-3.93 (m, CH₂, 1H), 3.77-3.70(m, CH₂, 1H), 3.54-3.43 (m, CH₂, 1H), 3.07-2.96 (m, CH₂, 1H), 2.93 (d,J=4.5 Hz, CH₃, 3H), 1.94 (br, CH₂, 2H), 1.85-1.80 (m, CH₂, 1H),1.48-1.46 (m, CH₂, 1H), 1.38 (s, CH₃×3, 9H); ESI-MS: m/z=504 [M+1]⁺.

Step 3. Synthesis ofN⁴-methyl-5-(4-methylthiazol-2-yl)-N²-(2-cyano-3-(piperidin-3-yloxy)pyridin-5-yl)-2,4-diaminopyrimidine(Compound 44)

Anhydrous DMF (10 mL) was added to a mixture of the compounds 2-45 (85.3mg, 0.211 mmol), 2-44 (124 mg, 0.422 mmol), Pd(dppf)Cl₂ (7.7 mg, 0.011mmol), CuCl (2.1 mg, 0.021 mmol), ZnCl₂ (28.8 mg, 0.211 mmol), Cs₂CO₃(137.5 mg, 0.422 mmol) under nitrogen atmosphere. The reaction washeated to 100° C. and stirred overnight. After suction filtration, thesolvent was removed under reduced pressure to give a residue, which waspurified by column chromatography on silica gel (eluentCH₂Cl₂:EtOH=30:1) to afford a white solid. The obtained white solid wasdissolved in dichloromethane (3 mL), trifluoroacetic acid (3 mL) wasadded dropwise, and the mixture was stirred for 30 minutes at ice-baththen stirred at room temperature for 4.5 hours. The mixture wasneutralized to pH 9 with saturated sodium hydrogen carbonate solution.Ethyl acetate (40 mL) was added, and the organic layer was washed withsaturated sodium chloride solution and dried over anhydrous sodiumsulfate. The organic layer was evaporated under reduced pressure and theresidue was purified by silica gel column chromatography, usingCH₂Cl₂/EtOH(NH₃) (100:3) as eluent to give a white solid. Yield: 40%; ¹HNMR (400 MHz, DMSO-d₆): δ 10.05 (s, NH, 1H), 8.46 (s, Ar—H, 1H), 8.41(s, Ar—H, 1H), 8.13 (s, Ar—H, 1H), 7.32 (q, J=4.5 Hz, NH, 1H), 4.59 (br,CH, 1H), 3.96-3.93 (m, CH₂, 1H), 3.77-3.70 (m, CH₂, 1H), 3.54-3.43 (m,CH₂, 1H), 3.07-2.96 (m, CH₂, 1H), 2.93 (d, J=4.5 Hz, CH₃, 3H), 1.94 (br,CH₂, 2H), 1.85-1.80 (m, CH₂, 1H), 1.48-1.46 (m, CH₂, 1H), 1.38 (s,CH₃×3, 9H); ESI-MS: m/z=504 [M+1]⁺.

Preparative Example 245-Trifluoromethyl-N-(2-cyano-3-(piperidin-3-oxy)pyridin-5-yl)-2-aminopyrimidine(Compound 45)

Step 1. Synthesis of 5-trifluoromethyl-2-aminopyrimidine (Intermediate2-47)

Compound 2-46 (225 mg, 1.23 mmol) was placed in a sealed tube, ammoniumhydroxide (10 mL) and N-methylpyrrolidone (10 mL) were added, and themixture was stirred at 120° C. for 24 h. After cooling to roomtemperature, the solvent was removed under reduced pressure to give aresidue. Purified by silica gel column chromatography (eluentPE:EtOAc=2:1) to give a white solid. LC-MS: m/z=164 [M+1]⁺.

Step 2. Synthesis of5-trifluoromethyl-N-(2-cyano-3-(piperidin-3-oxy)pyridin-5-yl)-2-aminopyrimidine(Compound 45)

Synthetic Procedure Reference Example 1, Step 5. Compound 45 wasprepared from intermediates 2-47 and 2-2 to give a white solid. LC-MS:m/z=365 [M+1]⁺.

Preparative Example 25. Preparation of Compound 46-50

Step 1. Synthesis of Intermediate 2-49/2-51

The synthetic steps were carried out according to the same procedure asin Example 17, Step 2, and Example 1, Step 2, respectively, to obtainthe compounds 2-49/2-51, and the mass spectral data were LC-MS: m/z=194[M+1]⁺ and LC-MS: m/z=193 [M+1]⁺, respectively.

Step 2. Synthesis of Intermediates 2-50/2-52

The synthetic steps were carried out according to the same procedure asin Example 24, Step 1. Compound 2-50/2-52 was prepared from 2-49/2-51,respectively, to give a white solid. The mass spectral data were LC-MS:m/z=194 [M+1]⁺ and LC-MS: m/z=193[M+1]⁺, respectively.

Step 3. Synthesis of Compound 46-50

The procedure of synthesis was described as that of Example 5, Step 5.Compound 46-50 were prepared from 2-16, 2-13, 2-35, 2-44 andintermediate 2-50/2-52, respectively, to give white solids. The massspectral data were LC-MS: m/z=409 [M+1]⁺, LC-MS: m/z=394 [M+1]⁺, LC-MS:m/z=394 [M+1]⁺, LC-MS: m/z=394 [M+1]⁺, and LC-MS: m/z=393 [M+1]⁺,respectively.

The NMR and MS data of the intermediates 2-21˜2-33, 2-3˜2-42, compounds19-31, and 34-43 are shown in Table 1-1, 1-2, 1-3, 1-4.

TABLE 1-1 Compounds Nuclear magnetic resonance and Number Name of theCompounds mass spectrometry data Intermediate 5-phenyl-N⁴-methyl-2, 4-¹H NMR (500 MHz, CDCl₃): δ 7.69 (br, Ar- 2-21 diaminopyrimidine H, 1H),7.49 (d, J = 5.5 Hz, Ar-H, 2H), 7.13- 7.09 (m, Ar-H, 3H), 6.43 (q, J =4.5 Hz, NH, 1H), 6.25 (br, NH2, 2H), 2.85 (d, J = 4.5 Hz, CH₃, 3H);ESI-MS: m/z = 201 [M + 1]⁺ Intermediate 5-(3-fluorophenyl)-N⁴- ¹H NMR(500 MHz, CDCl₃): δ 7.60 (br, Ar- 2-22 methyl-2, 4- H, 1H), 7.47-7.43(m, Ar-H, 1H), 7.16-7.11 diaminopyrimidine (m, Ar-H, 3H), 6.36 (q, J =4.5 Hz, NH, 1H), 6.21 (br, NH₂, 2H), 2.79 (d, J = 4.5 Hz, CH₃, 3H);ESI-MS: m/z = 219 [M + 1]⁺ Intermediate 5-(4-fluorophenyl)-N⁴- ¹H NMR(500 MHz, CDCl₃): δ 7.57 (s, Ar-H, 2-23 methyl-2, 4- 1H), 7.31 (d, J =8.5 Hz, Ar-H, 2H), 7.07 (d, diaminopyrimidine J = 8.5 Hz, Ar-H, 2H),6.17 (q, J = 4.5 Hz, NH, 1H), 6.11 (br, NH₂, 2H), 2.85 (d, J = 4.5 Hz,CH₃, 3H); ESI-MS: m/z = 219 [M + 1]⁺ Intermediate 5-(2-fluorophenyl)-N⁴-¹H NMR (500 MHz, CDCl₃): δ 7.49 (s, Ar-H, 2-24 methyl-2, 4- 1H),7.41-7.37 (m, Ar-H, 1H), 7.30- 7.27 (m, diaminopyrimidine Ar-H, 1H),7.25-7.22 (m, Ar-H, 2H), 6.08 (br, NH, 2H), 6.04 (q, J = 4.5 Hz, NH),2.75 (d, J = 4.5 Hz, CH₃, 3H); ESI-MS: m/z = 219 [M + 1]⁺ Intermediate5-(3 -methoxyphenyl)-N⁴ - ¹H NMR (500 MHz, DMSO-d₆): δ 7.55 (s, 2-25methyl-2, 4- Ar-H, 1H), 7.34-7.30 (m, Ar-H, 1H), 6.88-6.86diaminopyrimidine (m, Ar-H, 2H), 6.84 (s, Ar-H, 1H), 6.15 (q, J = 4.5Hz, NH), 6.01 (br, NH, , 2H), 3.78 (s, CH₃, 3H), 2.77 (d, J = 4.5 Hz,CH₃, 3H); ESI-MS: m/z = 231 [M + 1]⁺ Intermediate5-(4-methoxyphenyl)-N⁴- ¹H NMR (500 MHz, CDCl₃): δ 7.47 (_(s), Ar-H,2-26 methyl-2, 4- 1H), 7.22 (d, J = 8.5 Hz, Ar-H, 2H), 6.99 (d,diaminopyrimidine J = 8.5 Hz, Ar-H, 2H), 6.09 (q, J = 4.5 Hz, NH, 1H),6.03 (br, NH₂, 2H), 3.77 (s, CH₃, 3H), 2.76 (d, J = 4.5 Hz, CH₃, 3H);ESI-MS: m/z = 231 [M + 1]⁺ _(o) Intermediate 5 -(2, 4-dimethoxyphenyl) -¹H NMR (400 MHz, CDCl₃): δ 7.35 (br, Ar- 2-27 N⁴-methyl-2, 4- H, 1H),6.99 (d, J = 8.0 Hz, Ar-H, 1H), 6.61 diaminopyrimidine (s, Ar-H, 1H),6.57 (d, J = 8.0 Hz, Ar-H, 1H), 5.87 (br, NH₂, 2H), 5.62 (br, NH, 1H),3.79 (s, CH₃, 3H), 3.71 (s, CH₃, 3H), 2.72 (d, J = 4.5 Hz, CH₃, 3H);ESI-MS: m/z = 261 [M + 1]⁺ Intermediate 5-(pyri din-3-yl)-N⁴ - ¹H NMR(500 MHz, DMSO-d₆): δ 8.50 (br, 2-28 methyl-2, 4- Ar-H, 2H), 7.72 (d, J= 8.0 Hz, Ar-H, 1H), diaminopyrimidine 7.56 (s, Ar-H, 1H), 7.43-7.40 (m,Ar-H, 1H), 6.35 (q, J = 4.5 Hz, NH, 1H), 6.14 (br, NH, , 2H), 2.76 (d, J= 4.5 Hz, CH₃, 3H); ESI-MS: mz = 202 [M + 1]⁺ Intermediate5-(pyridin-4-yl)-N⁴- ¹H NMR (500 MHz, DMSO-d₆): δ 8.43 (br, 2-29methyl--2, 4- Ar-H, 1H), 7.31 (d, J = 8.5 Hz, Ar-H, 2H),diaminopyrimidine 7.08 (d, J = 8.5 Hz, Ar-H, 2H), 6.18 (q, J = 4.5 Hz,NH, 1H), 6.17 (br, NH₂, 2H), 2.82 (d, J = 4.5 Hz, CH₃, 3H); ESI-MS: m/z= 202 [M + 1]⁺ Intermediate 5-(thien-2-yl)-N⁴- ¹H NMR (500 MHz,DMSO-d₆): δ 7.66 (s, 2-30 methyl-2, 4- Ar-H, 1H), 7.51 (d, J = 4.5 Hz,Ar-H, 1H), diaminopyrimidine 7.13-7.11 (m, Ar-H, 1H), 7.06 (d, J = 2.5Hz, Ar-H, 1H), 6.30 (q, J = 4.5 Hz, NH, 1H), 6.21 (br, NH₂, 2H), 2.80(d, J = 4.5 Hz, CH₃, 3H): ESI-MS: m/z = 207 [M + 1]⁺ Intermediate5-(furan-2-yl)-N⁴- ¹H NMR (500 MHz, DMSO-d₆): δ 7.64 (s, 2-31 methyl-2,4- Ar-H, 1H), 7.57-7.48 (m, Ar-H, 1H), 6.54 (br, diaminopyrimidine Ar-H,2H), 6.44 (q, J = 3.5 Hz, NH, 1H), 6.22 (br, NH, 2H), 2.86 (d, J = 4.5Hz, CH₃, 3H); ESI-MS: m/z = 191 [M + 1]⁺ Intermediate 5-(5- ¹H NMR (500MHz, DMSO-d₆): δ 7.78 (d, J = 2-33 methoxycarbonylthien- 4.0 Hz, Ar-H,1H), 7.77 (s, Ar-H, 1H), 7.16 2-yl)-N⁴-methyl-2, 4- (d, J = 4.0 Hz,Ar-H, 1H), 6.56 (q, J = 4.5 Hz, diaminopyrimidine NH, 1H), 6.40 (br,NH₂, 2H), 3.82 (s, CH₃, 3H), 2.80 (d, J = 4.5 Hz, CH₃, 3H); ESI-MS: m/z= 265 [M + 1]⁺

TABLE 1-2 Compounds Nuclear magnetic resonance and Number Name of theCompounds mass spectrometry data Compound 19 N⁴-methyl-5-phenyl-N²- ¹HNMR (500 MHz, DMSO-d₆): δ 9.96 (2-cyano-3-(piperidin-3- (br, NH, 1H),8.54 (s, Ar-H, 1H), 8.53 (s, oxy)pyridin-5-yl)-2, 4- Ar-H, 1H), 7.82 (s,Ar-H, 1H), 7.50-7.47 (m, diaminopyrimidine Ar-H, 2H), 7.42-7.40 (m,Ar-H, 3H), 6.77 (q, J = 4.5 Hz, NH, 1H), 4.41-4.36 (m, CH, 1H), 3.16(dd, J = 12.0 Hz, 2.0 Hz, CH₂, 1H), 2.93 (d, J = 4.5 Hz, CH₃, 3H), 2.79-2.75 (m, CH₂, 1H), 2.64-2.60 (m, CH₂, 1H), 2.54-2.49 (m, CH₂, 1H),2.11-2.08 (m, CH₂, 1H), 1.75-1.69 (m, CH₂, 1H), 1.63-1.56 (m, CH₂, 1H),1.46-1.38 (m, CH₂, 1H); 13c NMR (125 MHz, DMSO-d₆): δ 160.20, 158.17,157.35, 153.93, 142.67, 134.67, 134.19, 129.02, 128.78, 127.46, 116.43,112.85, 112.77, 108.20, 74.26, 49.78, 45.27, 29.89, 28.28, 24.19;ESI-MS: m/z = 402 [M + 1]⁺ Compound 20 N⁴-methyl-5-(3- ¹H NMR (500 MHz,DMSO-d₆): δ 10.03 fluorophenyl)-N²-(2- (s, NH, 1H), 8.53 (d, J = 4.0 Hz,Ar-H, 2H), cyano-3-(piperidin-3 - 7.86 (s, Ar-H, 1H), 7.54 (q, J = 6.5Hz, Ar- yloxy)pyridin-5-yl)-2, 4- H, 1H), 7.26-7.21 (m, Ar-H, 3H), 6.92(q, diaminopyrimidine J = 4.0 Hz, NH, 1H), 4.41-4.38 (m, CH, 1H), 3.16(d, J = 10.5 Hz, CH₂, 1H), 2.92 (d, J = 4.5 Hz, CH₃, 3H), 2.78-2.76 (m,CH₂, 1H), 2.65-2.61 (m, CH₂, 1H), 2.55-2.53 (m, CH₂, 1H), 2.09 (br, CH₂,1H), 1.73-1.71 (m, CH₂, 1H), 1.63-1.56 (m, CH₂, 1H), 1.45-1.39 (m, CH₂,1H); ¹³C NMR (125 MHz, DMSO- d₆): δ 163.39, 161.45, 160.05, 158.36,157.29, 154.18, 142.58, 137.13, 137.07, 134.25, 130.95, 130.88, 124.97,116.40, 115.74, 115.57, 114.34, 114.17, 112.99, 111.62, 108.32, 74.07,49.50, 45.13, 29.71, 28.26, 23.88; ESI-MS: m/z = 420 [M + 1]⁺ Compound21 N⁴-methyl-5-(4- ¹H NMR (400 MHz, DMSO-d₆): δ 9.96fluorophenyl)-N²-(2- (br, NH, 1H), 8.53 (s, Ar-H, 1H), 8.52 (s, cyano-3-(piperidin-3 - Ar-H, 1H), 7.80 (s, Ar-H, 1H), 7.44-7.41 (m,yloxy)pyridin-5-yl)-2, 4- Ar-H, 2H), 7.31-7.28 (m, Ar-H, 2H), 6.77diaminopyrimidine (q, J = 4.6 Hz, NH, 1H), 4.40-4.35 (m, CH, 1H), 3.15(dd, J = 12.0 Hz, 2.0 Hz, CH₂, 1H), 2.92 (d, J = 3.6 Hz, CH₃, 3H), 2.78-2.74 (m, CH₂, 1H), 2.63-2.59 (m, CH₂, 1H), 2.54-2.49 (m, CH₂, 1H),2.11-2.07 (m, CH₂, 1H), 1.74-1.68 (m, CH₂, 1H), 1.63-1.56 (m, CH₂, 1H),1.45-1.37 (m, CH₂, 1H); 13c NMR (100 MHz, DMSO-d₆): δ 162.64, 160.70,160.32, 158.24, 157.34, 153.92, 142.64, 134.18, 131.08, 131.02, 116.41,115.89, 115.72, 112.87, 111.90, 108.22, 74.28, 49.80, 45.28, 29.89,28.22, 24.21; ESI-MS: mz = 420 [M + 1]⁺ Compound 22 N⁴-methyl-5-(2- ¹HNMR (400 MHz, DMSO-d₆) δ 9.99 (br, fluorophenyl)-N²-(2- NH, 1H), 8.55(s, Ar-H, 1H), 8.53 (s, Ar-H, cyano-3 -(piperidin-3 - 1H), 7.82 (s,Ar-H, 1H), 7.50-7.45 (m, Ar-H, oxy)pyridin-5-yl)-2, 4- 1H), 7.40-7.37(m, Ar-H, 1H), 7.33-7.29 (m, diaminopyrimidine Ar-H, 2H), 6.73 (q, J =3.6 Hz, NH, 1H), 4.42-4.37 (m, CH, 1H), 3.16 (dd, J = 9.6 Hz, 2.0 Hz,CH₂, 1H), 2.92 (d, J = 3.6 Hz, CH₃, 3H), 2.78-2.74 (m, CH₂, 1H),2.64-2.60 (m, CH₂, 1H), 2.54-2.49 (m, CH₂, 1H), 2.11- 2.08 (m, CH₂, 1H),1.73-1.70 (m, CH₂, 1H), 1.64-1.57 (m, CH₂, 1H), 1.46-1.38 (m, CH₂, 1H);¹³C NMR (100 MHz, DMSO-d₆): δ 160.92, 160.29, 158.97, 158.69, 157.33,154.77, 142.58, 134.24, 132.09, 130.10, 130.03, 124.92, 122.03, 121.90,116.38, 116.08, 115.91, 112.99, 108.40, 106.78, 74.30, 49.79, 45.28,29.88, 28.15, 24.20; ESI-MS: mz = 420 [M + 1]⁺ Compound 23N⁴-methyl-5-(3- ¹H NMR (400 MHz, DMSO-d₆): δ 9.96 methoxyphenyl)-N²-(2-(br, NH, 1H), 8.54 (s, Ar-H, 1H), 8.53 (s, cyano-3 -(piperidin-3 - Ar-H,1H), 7.84 (s, Ar-H, 1H), 7.40-7.37 (m, yloxy)pyridin-5-yl)-2, 4- Ar-H,1H), 6.97-6.95 (m, Ar-H, 3H), 6.79 diaminopyrimidine (q, J = 3.6 Hz, NH,1H), 4.40-4.35 (m, CH, 1H), 3.81 (s, CH₃, 3H), 3.16 (dd, J = 9.6 Hz, 1.6Hz, CH₂, 1H), 2.94 (d, J = 3.6 Hz, CH₃, 3H), 2.79-2.74 (m, CH₂, 1H),2.63-2.59 (m, CH₂, 1H), 2.54 (dd, J = 17.6 Hz, 2.4 Hz, CH₂, 1H),2.11-2.08 (m, CH₂, 1H), 1.74- 1.68 (m, CH₂, 1H), 1.63-1.56 (m, CH₂, 1H),1.46-1.37 (m, CH₂, 1H); ¹³C NMR (100 MHz, DMSO-d₆):8160.14, 159.60,158.16, 157.34, 153.81, 142.65, 135.97, 134.17, 130.06, 120.92, 116.41,114.06, 113.33, 112.85, 112.66, 108.19, 74.29, 55.01, 49.82, 45.29,29.91, 28.26, 24.23; ESI-MS: m/z = 432 [M + 1]⁺ Compound 24N⁴-methyl-5-(4- ¹H NMR (400 MHz, DMSO-d₆): δ 9.92 methoxyphenyl)-N²-(2-(s, NH, 1H), 8.53 (s, Ar-H, 2H), 7.77 (s, Ar- cyano-3 -(piperidin-3 - H,1H), 7.32 (d, J = 7.2 Hz, Ar-H, 2H), 7.05 yloxy)pyridin-5-yl)-2, 4- (d,J = 7.2 Hz, Ar-H, 2H), 6.68 (q, J = 3.6 diaminopyrimidine Hz, NH, 1H),4.40-4.35 (m, CH, 1H), 3.80 (s, CH₃, 3H), 3.15 (dd, J = 9.6 Hz, 1.6 Hz,CH₂, 1H), 2.92 (d, J = 3.6 Hz, CH₃, 3H), 2.78-2.74 (m, CH₂, 1H),2.63-2.59 (m, CH₂, 1H), 2.54-2.49 (m, CH₂, 1H), 2.11-2.07 (m, CH₂, 1H),1.74-1.68 (m, CH₂, 1H), 1.63- 1.56 (m, CH₂, 1H), 1.45-1.37 (m, CH₂, 1H);¹³C NMR (100 MHz, DMSO-d₆): δ 160.44, 158.74, 157.97, 157.35, 153.56,142.72, 134.14, 130.07, 126.64, 116.64, 114.48, 112.74, 112.59, 108.09,74.27, 55.15, 49.82, 45.29, 29.90, 28.25, 24.22; ESI-MS: m/z = 432 [M +1]⁺ Compound 25 N⁴-methyl-5-(2, 4- ¹H NMR (400 MHz, DMSO-d₆): δ 9.88dimethoxyphenyl)-N²- (s, NH, 1H), 8.55 (s, Ar-H, 1H), 8.53 (s, Ar-(2-cyano-3-(piperidin-3- H, 1H), 7.66 (s, Ar-H, 1H), 7.09 (d, J = 6.8yloxy)pyridin-5-yl)-2, 4- Hz, Ar-H, 1H), 6.67 (s, Ar-H, 1H), 6.62 (d,diaminopyrimidine J = 6.8 Hz, Ar-H, 1H), 6.33 (q, J = 3.6 Hz, NH, 1H),4.41-4.36 (m, CH, 1H), 3.82 (s, CH₃, 3H), 3.75 (s, CH₃, 3H), 3.16 (dd, J= 9.6 Hz, 1.6 Hz, CH₂, 1H), 2.90 (d, J = 3.6 Hz, CH₃, 3H), 2.79-2.74 (m,CH₂, 1H), 2.64- 2.60 (m, CH₂, 1H), 2.54-2.50 (m, CH₂, 1H), 2.11-2.08 (m,CH₂, 1H), 1.73-1.70 (m, CH₂, 1H), 1.63-1.56 (m, CH₂, 1H), 1.45-1.37 (m,CH₂, 1H); ¹³C NMR (100 MHz, DMSO- d₆): δ 160.74, 160.71, 158.25, 158.11,157.37, 154.11, 142.85, 134.14, 131.90, 116.48, 115.27, 112.61, 109.92,108.01, 105.31, 98.90, 74.20, 55.36, 55.27, 49.76, 45.25, 29.87, 28.16,24.15; ESI-MS: m/z = 462 [M + 1]⁺ Compound 26 N⁴-methyl-5-(pyridin-3- ¹HNMR (400 MHz, DMSO-d₆): δ 10.00 yl)-N²-(2-cyano-3- (br, NH, 1H),8.60-8.58 (m, Ar-H, 2H), 8.55 (piperidin-3- (s, Ar-H, 1H), 8.52 (s,Ar-H, 1H), 7.86 (s, yloxy)pyridin-5-yl)-2, 4- Ar-H, 1H), 7.83-7.81 (m,Ar-H, 1H), 7.50- diaminopyrimidine 7.47 (m, Ar-H, 1H), 6.98 (q, J = 3.6Hz, NH, 1H), 4.40-4.35 (m, CH, 1H), 3.16 (dd, J = 9.6 Hz, 2.0 Hz, CH₂,1H), 2.92 (d, J = 3.6 Hz, CH₃, 3H), 2.79-2.75 (m, CH₂, 1H), 2.64- 2.60(m, CH₂, 1H), 2.54-2.49 (m, CH₂, 1H), 2.11-2.08 (m, CH₂, 1H), 1.75-1.69(m, CH₂, 1H), 1.63-1.56 (m, CH₂, 1H), 1.46-1.38 (m, CH₂, 1H); ¹³CNMR(100 MHz, DMSO- de)-. 5 160.37, 158.58, 157.32, 154.50, 149.46,148.44, 142.54, 136.55, 134.22, 130.70, 123.84, 116.37, 113.01, 109.51,108.36, 74.31, 49.81, 45.29, 29.90, 28.20, 24.22; ESI-MS: m/z = 403 [M +1]⁺ Compound 27 N⁴-methyl-5-(pyridin-4- ¹H NMR (400 MHz, DMSO-d₆): δ10.06 yl)-N²-(2-cyano-3- (br, NH, 1H), 8.64 (d, J = 4.8 Hz, Ar-H,(piperidin-3 - 2H), 8.55 (s, Ar-H, 1H), 8.50 (s, Ar-H, 1H),yloxy)pyridin-5-yl)-2, 4- 7.94 (s, Ar-H, 1H), 7.46 (d, J = 4.8 Hz, Ar-diaminopyrimidine H, 2H), 7.06 (q, J = 3.6 Hz, NH, 1H), 4.41- 4.36 (m,CH, 1H), 3.16 (dd, J = 9.6 Hz, 2.0 Hz, CH₂, 1H), 2.94 (d, J = 3.6 Hz,CH₃, 3H), 2.79-2.75 (m, CH₂, 1H), 2.64-2.60 (m, CH₂, 1H), 2.55-2.51 (m,CH₂, 1H), 2.11-2.08 (m, CH₂, 1H), 1.73-1.71 (m, CH₂, 1H), 1.63- 1.56 (m,CH₂, 1H), 1.46-1.39 (m, CH₂, 1H); ¹³C NMR (100 MHz, DMSO-d₆): δ 160.30,159.20, 157.80, 155.17, 150.59, 143.14, 142.91, 134.78, 123.90, 116.85,113.68, 110.59, 109.00, 74.78, 50.26, 45.77, 30.37, 28.76, 24.68;ESI-MS: m/z = 403 [M + 1]⁺ Compound 28 N⁴-methyl-5-(thien-2- ¹H NMR (400MHz, DMSO-d₆): δ 10.05 yl)-N²-(2-cyano-3- (br, NH, 1H), 8.53 (s, Ar-H,1H), 8.51 (s, (piperidin-3 - Ar-H, 1H), 7.96 (s, Ar-H, 1H), 7.63 (d, J =oxy)pyridin-5-yl)-2, 4- 4.0 Hz, Ar-H, 1H), 7.22-7.19 (m, Ar-H, 2H),diaminopyrimidine 6.95 (q, J = 3.6 Hz, NH, 1H), 4.41-4.37 (m, CH, 1H),3.16 (dd, J = 9.6 Hz, 2.0 Hz, CH₂, 1H), 2.97 (d, J = 3.6 Hz, CH₃, 3H),2.79- 2.75 (m, CH₂, 1H), 2.63-2.59 (m, CH₂, 1H), 2.54-2.49 (m, CH₂, 1H),2.11-2.08 (m, CH₂, 1H), 1.73-1.70 (m, CH₂, 1H), 1.63-1.56 (m, CH₂, 1H),1.46-1.38 (m, CH₂, 1H); ¹³C NMR (100 MHz, DMSO-d₆): δ 160.12, 158.32,157.34, 154.46, 142.45, 135.46, 134.25, 128.11, 126.62, 126.02, 116.40,113.09, 108.40, 105.78, 74.29, 49.83, 45.32, 29.93, 28.39, 24.26;ESI-MS: m/z = 408 [M + 1]⁺ Compound 29 N⁴-methyl-5-(furan-2- ¹H NMR (400MHz, DMSO-d₆): δ 10.07 yl)-N²-(2-cyano-3- (br, NH, 1H), 8.53 (d, J = 1.6Hz, Ar-H, (piperidin-3- 1H), 8.48 (d, J = 1.6 Hz, Ar-H, 1H), 8.25 (s,yloxy)pyridin-5-yl)-2, 4- Ar-H, 1H), 7.75 (s, Ar-H, 1H), 7.07 (q, J =diaminopyrimidine 3.6 Hz, NH, 1H), 6.77 (d, J = 2.8 Hz, Ar-H, 1H),6.63-6.62 (m, Ar-H, 1H), 4.41-4.37 (m, CH, 1H), 3.16 (dd, J = 9.6 Hz,2.0 Hz, CH₂, 1H), 3.04 (d, J = 3.6 Hz, CH₃, 3H), 2.80- 2.76 (m, CH₂,1H), 2.64-2.60 (m, CH₂, 1H), 2.55-2.53 (m, CH₂, 1H), 2.11-2.08 (m, CH₂,1H), 1.75-1.69 (m, CH₂, 1H), 1.64-1.57 (m, CH₂, 1H), 1.47-1.39 (m, CH₂,1H); ESI- MS: m/z = 392 [M + 1]⁺ Compound 30 N⁴-methyl-5-(5-chloro- ¹HNMR (400 MHz, DMSO-d₆): δ 10.07 furan-2-yl)-N²-(2-cyano- (br, NH, 1H),8.53 (d, J = 1.6 Hz, Ar-H, 3-(piperidin-3- 1H), 8.48 (d, J = 1.6 Hz,Ar-H, 1H), 8.25 (s, yloxy)pyridin-5-yl)-2, 4- Ar-H, 1H), 7.75 (s, Ar-H,1H), 7.07 (q, J = diaminopyrimidine 3.6 Hz, NH, 1H), 6.77 (d, J = 2.8Hz, Ar-H, 1H), 6.63-6.62 (m, Ar-H, 1H), 4.41-4.37 (m, CH, 1H), 3.16 (dd,J = 9.6 Hz, 2.0 Hz, CH₂, 1H), 3.04 (d, J = 3.6 Hz, CH₃, 3H), 2.80- 2.76(m, CH₂, 1H), 2.64-2.60 (m, CH₂, 1H), 2.55-2.53 (m, CH₂, 1H), 2.11-2.08(m, CH₂, 1H), 1.75-1.69 (m, CH₂, 1H), 1.64-1.57 (m, CH₂, 1H), 1.47-1.39(m, CH₂, 1H); ESI- MS: mz = 426 [M + 1]⁺ Compound 31 N⁴-methyl-5-(5- ¹HNMR (400 MHz, DMSO-d₆): δ 10.10 methoxycarbonylthien- (br, NH, 1H), 8.53(s, Ar-H, 1H), 8.46 (s, 2-yl)-N²-(2-cyano-3- Ar-H, 1H), 8.03 (s, Ar-H,1H), 7.83 (d, J = (piperidin-3- 3.2 Hz, Ar-H, 1H), 7.29 (d, J = 2.8 Hz,Ar- yloxy)pyridin-5-yl)-2, 4- H, 1H), 7.12 (q, J = 3.2 Hz, NH, 1H),4.39- diaminopyrimidine 4.36 (m, CH, 1H), 3.86 (s, CH₃, 3H), 3.17 (dd, J= 9.6 Hz, 1.6 Hz, CH₂, 2H), 2.97 (d, J = 3.2 Hz, CH₃, 3H), 2.80-2.77 (m,CH₂, 1H), 2.65-2.61 (m, CH₂, 1H), 2.10-2.08 (m, CH₂, 1H), 1.74-1.71 (m,CH₂, 1H), 1.64-1.57 (m, CH₂, 1H), 1.47-1.39 (m, CH₂, 1H); ¹³C NMR (100MHz, DMSO-d₆): δ 161.67, 159.83, 158.71, 157.26, 155.02, 143.51, 142.20,134.40, 131.32, 127.40, 116.34, 113.37, 108.61, 104.83, 74.25, 52.23,49.76, 45.30, 29.88, 28.40, 24.17. ESI-MS: m/z = 466 [M + 1]⁺

TABLE 1-3 Compounds Nuclear magnetic resonance and Number Name of theCompounds mass spectrometry data Intermediate 2-36(R)-2-cyano-5-bromo-3-(N- ¹H NMR (500 MHz, DMSO-d₆): δtert-butoxycarbonylpyrrole- 8.48 (d, J = 2.0 Hz, Ar-H, 1H), 8.29 (s,3-yloxy)pyridine Ar-H, 1H), 5.32 (br, CH, 1H), 3.62- 3.55 (m, CH, 1H),3.49-3.44 (m, CH₂, 2H), 3.39-3.33 (m, CH₂, 1H), 2.21- 2.16 (m, CH₂, 1H),2.13-2.09 (m, CH₂, 1H), 1.41 (s, CH₃ × 3, 9H) ; ESI- MS: m/z = 368 [M +1]⁺ Intermediate 2-37 (S)-2-cyano-5-bromo-3-(N- ¹H NMR (500 MHz,DMSO-d₆): δ tert-butoxycarbonylpyrrole- 8.48 (d, J = 2.0 Hz, Ar-H, 1H),8.29 (s, 3-yloxy)pyridine Ar-H, 1H), 5.32 (br, CH, 1H), 3.62- 3.55 (m,CH, 1H), 3.49-3.44 (m, CH₂, 2H), 3.39-3.33 (m, CH₂, 1H), 2.21- 2.16 (m,CH₂, 1H), 2.13-2.09 (m, CH₂, 1H), 1.41 (s, CH₃ × 3, 9H) ; ESI- MS: m/z =368 [M + 1]⁺ Intermediate 2-38 2-cyano-5-bromo-3-(N-tert- ¹H NMR (500MHz, CDCl₃): δ 8.34 butoxycarbonylpiperidin-4- (s, Ar-H, 1H), 7.51 (s,Ar-H, 1H), yloxy)pyridine 4.66-4.62 (m, CH, 1H), 3.69-3.63 (m, CH₂, 2H),3.52-3.46 (m, CH₂, 2H), 1.99-1.92 (m, CH₂, 2H), 1.90-1.82 (m, CH₂, 2H),1.47 (s, CH₃ × 3, 9H); ESI- MS: m/z = 382 [M + 1]⁺ Intermediate 2-392-cyano-5-bromo-3-(2- ¹H NMR (500 MHz, CDCl₃): δ 8.27dimethylamino)ethoxypyridine (s, Ar-H, 1H), 7.49 (s, Ar-H, 1H), 4.16 (t,J = 7.0 Hz, CH₂, 2H), 2.78 (t, J = 7.0 Hz, CH₂, 2H), 2.31 (s, CH₃ × 2,6H); ESI-MS: m/z = 270 [M + 1]⁺ Intermediate 2-40(R)-2-cyano-5-bromo-3-(1- ¹H NMR (500 MHz, CDCl₃): δ 8.31(dimethylamino)propyl-2- (s, Ar-H, 1H), 7.62 (s, Ar-H, 1H), oxy)pyridine4.62-4.55 (m, CH, 1H), 2.76-2.71 (m, CH₂, 1H), 2.54-2.49 (m, CH₂, 1H),2.31 (s, CH₃ × 2, 6H), 1.40 (d, J = 8.0 Hz, CH₃, 3H); ESI-MS: m/z = 284[M + 1]⁺ Intermediate 2-41 (S)-2-cyano-5-bromo-3-(1 - ¹H NMR (500 MHz,CDCl₃): δ 8.31 (dimethylamino)propyl-2- (s, Ar-H, 1H), 7.62 (s, Ar-H,1H), oxy)pyridine 4.62-4.55 (m, CH, 1H), 2.76-2.71 (m, CH₂, 1H),2.54-2.49 (m, CH₂, 1H), 2.31 (s, CH₃ × 2, 6H), 1.40 (d, J = 8.0 Hz, CH₃,3H); ESI-MS: m/z = 284 [M + 1]⁺

TABLE 1-4 Compounds Name of the Nuclear magnetic resonance and NumberCompounds mass spectrometry data Compound 34 N⁴-methyl-5-(1-methyl- ¹HNMR (400 MHz, DMSO-d₆): δ 9.96 1H-pyrazol-4-yl)-N²-(2- (br, NH, 1H),8.60 (s, Ar-H, 1H), 8.42 (s, cyano-3-(piperidin-4- Ar-H, 1H), 7.90 (s,Ar-H, 1H), 7.89 (s, Ar- methyl)oxypyridin-5-yl)- H, 1H), 7.62 (s, Ar-H,1H), 6.71 (q, J = 4.4 2,4-diaminopyrimidine Hz, NH, 1H), 3.95 (d, J =6.4 Hz, CH₂, 2H), 3.89 (s, CH₃, 3H), 2.99-2.96 (m, CH₂, 2H), 2.94 (d, J= 4.4 Hz, CH₃, 3H), 2.52-2.47 (m, CH₂, 2H), 1.89-1.86 (m, CH, 1H),1.71-1.68 (m, CH₂, 2H), 1.22-1.12 (m, CH₂, 2H); ESI-MS: m/z = 420 [M +1]⁺ Compound 35 (R)-N⁴-methyl-5-(1- ¹H NMR (400 MHz, DMSO-d₆): δ 9.91methyl-1H-pyrazol-4-yl)- (s, NH, 1H), 8.52 (s, Ar-H, 1H), 8.51 (s, Ar-N²-(2-cyano-3-(piperidin- H, 1H), 7.91 (s, Ar-H, 1H), 7.90 (s, Ar-H,3-yloxy)pyridin-5-yl)-2,4- 1H), 7.61 (s, Ar-H, 1H), 6.73 (q, J = 4.0 Hz,diaminopyrimidine NH, 1H), 4.40-4.35 (m, CH, 1H), 3.89 (s, CH₃, 3H),3.15 (dd, J = 9.6 Hz, 2.0 Hz, CH₂, 1H), 2.95 (d, J = 3.6 Hz, CH₃, 3H),2.79-2.75 (m, CH₂, 1H), 2.64-2.60 (m, CH₂, 1H), 2.54-2.51 (m, CH₂, 1H),2.11-2.07 (m, CH₂, 1H), 1.75-1.69 (m, CH₂, 1H), 1.63- 1.56 (m, CH₂, 1H),1.46-1.38 (m, CH₂, 1H); ¹³C NMR (100 MHz, DMSO-d₆): δ 160.27, 157.75,157.37, 153.08, 142.73, 137.65, 134.13, 129.42, 116.48, 113.78, 112.70,108.03, 104.51, 74.25, 49.80, 45.30, 38.65, 29.90, 28.28, 24.20; ESI-MS:m/z = 406 [M + 1]⁺ Compound 36 (S)-N⁴-methyl-5-(1- ¹H NMR (400 MHz,DMSO-d₆): δ 9.91 methyl-1H-pyrazol-4-yl)- (s, NH, 1H), 8.52 (s, Ar-H,1H), 8.51 (s, Ar- N²-(2-cyano-3-(piperidin- H, 1H), 7.91 (s, Ar-H, 1H),7.90 (s, Ar-H, 3-yloxy)pyridin-5-yl)-2,4- 1H), 7.61 (s, Ar-H, 1H), 6.73(q, J = 4.0 Hz, diaminopyrimidine NH, 1H), 4.40-4.35 (m, CH, 1H), 3.89(s, CH₃, 3H), 3.15 (dd, J = 9.6 Hz, 2.0 Hz, CH₂, 1H), 2.95 (d, J = 3.6Hz, CH₃, 3H), 2.79-2.75 (m, CH₂, 1H), 2.64-2.60 (m, CH₂, 1H), 2.54-2.51(m, CH₂, 1H), 2.11-2.07 (m, CH₂, 1H), 1.75-1.69 (m, CH₂, 1H), 1.63- 1.56(m, CH₂, 1H), 1.46-1.38 (m, CH₂, 1H); ¹³C NMR (100 MHz, DMSO-d₆): δ160.27, 157.75, 157.37, 153.08, 142.73, 137.65, 134.13, 129.42, 116.48,113.78, 112.70, 108.03, 104.51, 74.25, 49.80, 45.30, 38.65, 29.90,28.28, 24.20; ESI-MS: m/z = 406 [M + 1]⁺ Compound 37 (R)-N⁴-methyl-5-(1-¹H NMR (400 MHz, DMSO-d₆): δ 9.92 methyl-1H-pyrazol-4-yl) - (br, NH,1H), 8.60 (s, Ar-H, 1H), 8.39 (s, N²-(2-cyano-3-(pyrrole-3- Ar-H, 1H),7.91 (s, Ar-H, 2H), 7.62 (s, Ar- oxy)pyridin-5-yl)-2,4- H, 1H), 6.72 (q,J = 4.4 Hz, NH, 1H), 4.96 diaminopyrimidine (br, CH, 1H), 3.89 (s, CH₃,3H), 3.16 (dd, J = 12.4 Hz, 5.2 Hz, CH₂, 1H), 2.94-2.93 (m, CH₂, 2H),2.92 (d, J = 4.4 Hz, CH₃, 3H), 2.83-2.79 (m, CH₂, 1H), 2.10-2.02 (m,CH₂, 1H), 1.87-1.84 (m, CH₂, 1H); ¹³C NMR (100 MHz, DMSO-d₆): δ 160.28,157.72, 157.58, 152.97, 142.66, 137.65, 133.79, 129.43, 116.48, 113.76,112.41, 107.91, 104.64, 80.01, 52.80, 45.48, 38.66, 33.25, 28.15;ESI-MS: m/z = 392 [M + 1]⁺ Compound 38 (S)-N⁴-methyl-5-(1- ¹H NMR (400MHz, DMSO-d₆): δ 9.92 methyl-1H-pyrazol-4-yl)- (br, NH, 1H), 8.60 (s,Ar-H, 1H), 8.39 (s, N²-(2-cyano-3-(pyrrole-3- Ar-H, 1H), 7.91 (s, Ar-H,2H), 7.62 (s, Ar- oxy)pyridin-5-yl)-2,4- H, 1H), 6.72 (q, J = 4.4 Hz,NH, 1H), 4.96 diaminopyrimidine (br, CH, 1H), 3.89 (s, CH₃, 3H), 3.16(dd, J = 12.4 Hz, 5.2 Hz, CH₂, 1H), 2.94-2.93 (m, CH₂, 2H), 2.92 (d, J =4.4 Hz, CH₃, 3H), 2.83-2.79 (m, CH₂, 1H), 2.10-2.02 (m, CH₂, 1H),1.87-1.84 (m, CH₂, 1H); ESI-MS: m/z = 392 [M + 1]⁺ Compound 39N⁴-methyl-5-(1-methyl- ¹H NMR (400 MHz, DMSO-d₆): δ 9.901H-pyrazol-4-yl)-N²-(2- (s, NH, 1H), 8.53 (s, Ar-H, 1H), 8.47 (s, Ar-cyano-3-(piperidin-4- H, 1H), 7.91 (s, Ar-H, 1H), 7.90 (s, Ar-H,yloxy)pyridin-5-yl)-2,4- 1H), 7.61 (s, Ar-H, 1H), 6.73 (q, J = 4.8 Hz,diaminopyrimidine NH, 1H), 4.58-4.54 (m, CH, 1H), 3.88 (s, CH₃, 3H),3.00-2.96 (m, CH₂, 2H), 2.94 (d, J = 4.8 Hz, CH₃, 3H), 2.58-2.53 (m,CH₂, 2H), 1.96-1.92 (m, CH₂, 2H), 1.61-1.53 (m, CH₂, 2H); ESI-MS: m/z =406 [M + 1]⁺ Compound 40 N⁴-methyl-5-(1-methyl- ¹H NMR (400 MHz,DMSO-d₆): δ 9.96 1H-pyrazol-4-yl)-N²-(2- (s, NH, 1H), 8.56 (s, Ar-H,1H), 8.48 (s, Ar- cyano-3-(2- H, 1H), 7.91 (s, Ar-H, 1H), 7.90 (s, Ar-H,dimethylaminoethoxy) 1H), 7.61 (s, Ar-H, 1H), 6.73 (q, J = 4.4 Hz,pyridin-5-yl)-2,4- NH, 1H), 4.24 (t, J = 5.6 Hz, CH₂, 2H), 3.88diaminopyrimidine (s, CH₃, 3H), 2.94 (d, J = 4.8 Hz, CH₃, 3H), 2.72 (t,J = 5.6 Hz, CH₂, 2H), 2.25 (s, CH₃ × 2, 6H) ; ¹³C NMR(100 MHz, DMSO-d) :δ 160.22, 158.43, 157.73, 153.05, 142.82, 137.60, 133.83, 129.37,116.42, 113.77, 111.72, 106.92, 104.53, 66.99, 57.06, 45.54, 38.63,28.17; ESI- MS: mz = 394 [M + 1]⁺ Compound 41 (R)-N⁴-methyl-5-(1- ¹H NMR(400 MHz, DMSO-d₆): δ 9.94 methyl-1H-pyrazol-4-yl) - (s, NH, 1H), 8.54(s, Ar-H, 2H), 7.92 (s, Ar- N²-(2-cyano-3-(1- H, 1H), 7.91 (s, Ar-H,1H), 7.63 (s, Ar-H, dimethylaminopropyl-2- 1H), 6.74 (q, J = 4.4 Hz, NH,1H), 4.69- oxy)pyridin-5-yl)-2, 4- 4.65 (m, CH, 1H), 3.91 (s, CH₃, 3H),2.96 diaminopyrimidine (d, J = 4.4 Hz, CH₃, 3H), 2.63-2.58 (m, CH₂, 1H),2.53-2.48 (m, CH₂, 1H), 2.24 (s, CH₃ × 2, 6H), 1.35 (d, J = 6.0 Hz, CH₃,3H); ESI-MS: m/z = 408 [M + 1]⁺ Compound 42 (S)-N⁴-methyl-5-(1- ¹H NMR(400 MHz, DMSO-d₆): δ 9.94 methyl-1H-pyrazol-4-yl) - (s, NH, 1H), 8.54(s, Ar-H, 2H), 7.92 (s, Ar- N²-(2-cyano-3-(1- H, 1H), 7.90 (s, Ar-H,1H), 7.63 (s, Ar-H, dimethylaminopropyl-2- 1H), 6.73 (q, J = 4.4 Hz, NH,1H), 4.69- oxy)pyridin-5-yl)-2,4- 4.65 (m, CH, 1H), 3.90 (s, CH₃, 3H),2.95 diaminopyrimidine (d, J = 4.4 Hz, CH₃, 3H), 2.62-2.58 (m, CH₂, 1H),2.51-2.48 (m, CH₂, 1H), 2.24 (s, CH₃ × 2, 6H), 1.35 (d, J = 6.0 Hz, CH₃,3H); ESI-MS: m/z = 408 [M + 1]⁺ Compound 43 N⁴-methyl-5-(1-methyl- ¹HNMR (400 MHz, DMSO-d₆): δ 9.91 1H-pyrazol-4-yl)-N²-(2- (s, NH, 1H), 8.53(s, Ar-H, 1H), 8.47 (s, Ar- cyano-3-(N- H, 1H), 7.91 (s, Ar-H, 1H), 7.90(s, Ar-H, methylpiperidin-4- 1H), 7.62 (s, Ar-H, 1H), 6.73 (q, J = 4.4Hz, yloxy)pyridin-5-yl-2,4- NH, 1H), 4.54 (br, CH, 1H), 3.89 (s, CH₃,diaminopyrimidine 3H), 2.94 (d, J = 4.0 Hz, CH₃, 3H), 2.58 (br, CH₂,2H), 2.22 (br, CH₂, 2H), 2.19 (s, CH₃, 3H), 1.97 (br, CH₂, 2H), 1.77(br, CH₂, 2H); ESI-MS: mz = 420 [M + 1]⁺

Chk1 Inhibition of the Compounds Disclosed in the Present Invention

With the Saurosporine as a positive control, the Chk1 enzyme inhibitoryactivity (IC₅₀) was evaluated using the ADP-Glo kit. The compound actson Chk1 protein kinase and inhibits its phosphorylation substrateCdc25C. The phosphorylation process consumes ATP. After the reaction,ADP-Glo™ Reagent consumes the remaining ATP. The ADP produced during thereaction can be transformed by ADP-Glo Detection Reagent. For ATP, ATPacts as a substrate for the Ultra-Glo™ luciferase catalytic reaction,producing an optical signal. The test compound was dissolved in DMSO tomake a 10 mM stock solution and diluted to 12 different concentrationsin a certain ratio for testing. In a 384-well plate, add 1 μL of thetest compound to each well, 2 μL of 2.5× Chk1 kinase, add 2 μL of 1×buffer to the control group, incubate for 10 min at room temperature,and add 2 μL of 2.5× substrate at 37° C. Incubate for 1 h, stop thereaction by adding 5 μL of ADP-Glo™ Reagent, and incubate for 1 h at 37°C. 10 μL of ADP-Glo Detection Reagent was added, incubated at 37° C. for30 min, and three parallel wells were set for each sample. Absorbancewas measured by luminescence fluorescence microplate reader, and datawere calculated using GraphPad Prism 5 software to calculate IC₅₀values.

Inhibitory Activity of the Compounds Disclosed in the Present Inventionon Chk1 Kinase

TABLE 2 IC₅₀ (μM) of compounds against Chk1 kinase Chk1 Chk1 Compd.(IC₅₀, nM) Compd. (IC₅₀, μM) 1 >10 μM 24 6 2 >10 μM 25 12 3 >10 μM 269.5 4 >10 μM 27 78.5 5 >10 μM 28 9.2 6 >10 μM 29 4.7 7 >10 μM 30 387.78 >10 μM 31 14.2 9 4.7 32 6.9 10 16 33 4.8 11 14 34 0.6 12 7.8 35 6.8 135.1 36 5.3 14 49 37 1.0 15 25 38 893.8 16 1.9 39 17.0 17 8.8 40 0.4 181.8 41 8.6 19 50 42 1.1 20 >10 μM 43 12 21 >10 μM 44 0.7 22 >10 μM 45 <1μM 23 >10 μM 46 <1 μM 47  <1 μM 48 <1 μM 49  <1 μM 50 <1 μMStaurosporine 1.2

As can be seen from the data in the table, most of the compounds arepotent inhibitors of Chk1 protein kinase. The Chk1 inhibitory activityof 24 compounds is comparable to that of the positive compoundStaurosporine, and the 5 compounds are superior to the positive controlStaurosporine. Therefore, the 2-substituted pyrimidine derivatives usedas Chk1 inhibitors of the present invention have broad antitumorapplication prospects.

Proliferative Inhibitory Activity of the Compounds Disclosed in thePresent Invention on Various Tumor Cells

Cell lines: human multiple myeloma cells RPMI 8226, human mantle celllymphoma cells Mino, Jeko-1, human lymphoma cells Romas, human acutemonocytic leukemia cells MV-4-11, human breast cancer cells MCF-7 Humanlung cancer cell A549, human prostate cancer cell LnCAP, human gastriccancer cell BGC-823, human colon cancer cell HCT116, Colo205, humanovarian cancer cell OVCAR-8 Experimental method: MTS assay for in vitroproliferation of compounds against different tumor cell lines Inhibitoryactivity (IC₅₀).

The cells in the logarithmic growth phase were trypsinized, counted, andseeded at a density of 1×10⁴ cells/well in a 96-well plate at 100 μL perwell in a 37° C. incubator containing 5% CO₂ overnight. For the culture,six concentration gradients were set for each compound, and three setsof duplicate wells were set for each concentration. After the addition,the cells were cultured for 72 hours, and 20 μL of MTS was added. Afterincubating for 2 hours at 37° C., the absorbance at 490 nm (L1) wasmeasured with a SpectraMAX 340 microplate reader. The referencewavelength was 690 nm (L2), and the (L1-L2) value was plotted againstthe different concentrations of the inhibitor, half of the inhibitoryconcentration IC₅₀ was fitted by the formula.

TABLE 3-1 Inhibition of proliferative of compounds on each tumor cellline IC₅₀(μM) Cpd. RPMI8226 Mino Romas Jeko-1 MV-4-11 9 3.339 0.7080.536 0.342 0.044 12 3.597 0.608 0.401 0.253 0.035 13 NT^(b) NT^(b)NT^(b) NT^(b) 0.035 16 NT^(b) NT^(b) NT^(b) NT^(b) 0.107 17 3.290 0.4950.348 0.117 0.050 18 NT^(b) NT^(b) NT^(b) NT^(b) 0.036 29 2.543 0.1570.126 0.039 0.039 32 8.175 0.859 0.789 0.189 0.064 33 NT^(b) NT^(b)NT^(b) NT^(b) 0.040 34 NT^(b) NT^(b) NT^(b) NT^(b) 0.044 35 1.273 0.6470.356 0.339 0.053 36 2.493 0.956 0.649 0.548 0.072 37 1.578 0.419 0.2920.103 0.022 40 3.781 0.923 0.874 0.309 0.101 41 0.945 0.473 0.119 0.0940.034 42 0.814 0.227 0.218 0.137 0.092 44 0.448 0.128 0.117 0.084 0.023

TABLE 3-2 IC₅₀(μM)^(a) Cpd. MCF-7 A549 LnCAP BGC-823 HCT116 OVCAR-8Colo205 9 1.234 3.714 9.145 >10 5.217 0.513 2.843 12 2.541 2.962 7.8609.10.2 4.110 0.274 3.741 13 NT^(b) 1.843 6.779 8.130 4.556 0.662 0.94716 NT^(b) 3.474 8.776 >10 3.990 1.527 2.013 17 NT^(b) 4.238 >10 >101.896 0.220 0.878 18 NT^(b) 3.172 6.998 7.113 1.774 2.136 1.009 29 2.1782.183 >10 >10 1.225 0.329 0.624 32 NT^(b) 5.170 5.996 7.642 4.183 NT^(b)NT^(b) 33 NT^(b) 3.165 5.102 6.183 2.845 NT^(b) NT^(b) 34 NT^(b) 3.1924.628 >10 1.779 2.110 3.760 35 1.008 2.162 2.173 4.810 0.980 0.789 0.66336 NT^(b) 5.741 4.892 NT^(b) 2.547 4.178 2.180 37 0.945 1.784 0.9213.175 0.884 1.230 3.147 40 NT^(b) 3.512 7.164 NT^(b) 2.184 2.942 5.16041 1.009 2.173 2.620 5.170 1.312 1.187 0.993 42 NT^(b) >10 5.742 NT^(b)4.189 2.174 1.032 44 1.134 3.761 2.146 3.569 1.208 5.784 2.177 ^(a)IC₅₀:average of three experiments; ^(b)not tested.

The Activity of the Compound Disclosed in the Present Invention inCombination with Other Drugs

MV 4-11 cells were seeded at 5000/well into 96-well plates. When used incombination, the drug is determined according to the ratio of IC₅₀ ofthe two drugs. The concentration range of each drug is IC₂₀˜IC₈₀ (or ⅛,¼, ½, 1, 2 and 4 of IC₅₀). After 72 hours, cell viability was measuredby the addition of MTS reagent, and the inhibition rate Fa wascalculated as 100% of the unmedicated group. The Chou-Talalay method wasused to analyze the inhibition rate Fa and the corresponding drugconcentration into CompuSyn software, and the CI value and Fa-CI curveof single concentration drug were obtained. CI (combination index) iscalculated as CI=DA/ICX, A+DB/ICX, B (A, B stands for two differentdrugs, ICX, A and ICX, B is the growth inhibition rate when the twodrugs are used alone. The drug concentration at X, DA and DB are theconcentrations of the two drugs when the growth inhibition rate reachesX when the two drugs are combined. The results are shown in FIG. 1 . Inthe FIGURE: CHK1 inhibitor (35); FLT3 inhibitor Crenolanib (Cre),Quizartinib (Qui); Akt inhibitor GSK2141795 (GSK); CI=combination index,according to the judgment method of Soriano et al, 0.9≤CI≤1.1Superposition effect, 0.8≤CI<0.9 is a low degree synergy, 0.6≤CI<0.8 isa moderate synergy, 0.4≤CI<0.6 is a highly synergistic effect, and0.2≤CI<4 is a strong synergistic effect.

What is claimed is:
 1. A 2-polysubstituted aromatic ring-pyrimidine derivative, which is selected from the group consisting of: N⁴-methyl-5-(1-methyl-1H-pyrazol-4-yl)-N²-(3-(piperidin-4-yloxy)isoxazol-5-yl)-2,4-diaminopyrimidine. 